Protective Effects of N-6 Fatty Acids-Enriched Diet on Intestinal Ischaemia/Reperfusion Injury Involve Lipoxin A4 and Its Recept

Protective effects of n-6 fatty acids-enriched diet on intestinal ischaemia/reperfusion injury involve lipoxin A4 and its receptor Thomas Gobbetti, Simon Ducheix, Pauline Le Faouder, Teresa Pérez-Berezo, Fabien Riols, Jérôme Boué, Justine Bertrand-Michel, Marc Dubourdeau, Hervé Guillou, Mauro Perretti, et al.

To cite this version:

Thomas Gobbetti, Simon Ducheix, Pauline Le Faouder, Teresa Pérez-Berezo, Fabien Riols, et al.. Protective effects of n-6 fatty acids-enriched diet on intestinal ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, Wiley, 2015, 172 (3), pp.910-923. 10.1111/bph.12957. hal-01601458

HAL Id: hal-01601458 https://hal.archives-ouvertes.fr/hal-01601458 Submitted on 27 May 2020

HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 22 21 20 19 18 17 16 15 14 13 12 11 10 3 2 1 9 8 7 6 5 4

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, This article isprotectedbycopyright. Allrights reserved. 10.1111/bph.12957 as doi: article this cite Please ofRecord. the and Version version this between differences to lead maywhich process, proofreading and pagination typesetting, copyediting, the through not been has review but full peer undergone and publication for hasaccepted been article This +33 562744558 +33 562744500 31024 Toulouse,France CHU Purpan, BP3028 InsermBat.B U1043, Dr. Vergnolle Nathalie authors:Corresponding 8 Toulouse, France 7 6 5 4 Purpan Toulouse,France (CPTP), 3 2 1 Michel T Gobbetti title:Running Inserm France U1048.Toulouse, Inserm, France U1043,Toulouse, Protective effects of n-6 fatty acids-enriched diet on intestinal ischemia/reperfusion injury Protective acids-enrichedinjury ischemia/reperfusion intestinal fatty dieton n-6 effectsof LipidomicFacility, Metatoul Corede Platform,PaulSabatier, Université Toulouse, ToxalimUMR1331, Toulouse France WHRI,London, University, Queen Mary UK Université Université de PaulSabatier,Centre Toulouse, de Physiopathologie Toulouse de U5282,Toulouse,CNRS, France Ambiotis Ambiotis Toulouse,SAS, France N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal 6,7 , MDubourdeau

Accepted Article1,2,3,4 , S Ducheix, S Effects of n Effects of

910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument: 8 , HGuillou - 5 6 dietonischemia/reperfusion injury , P leFaouder, P involve lipoxin A4 and lipoxinA4and involve its receptor.

5 , MPerretti 1,2,3,6,7 ,

T 4 Perez , NVergnolle 1,2,3 , F Riols

1,2,3 1

6,7 , NCenac , BoueJ

1,2,3 1,2,3

, Bertrand-J Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 24 23

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument:

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 31 30 29 28 27 26 25 42 41 40 39 38 37 36 35 34 33 32

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, Abbreviations: Abbreviations: This article isprotectedbycopyright. Allrights reserved. B thromboxane (PDx), (PGE A prostaglandin (PUFA), acids (MPO), ( acid oxoeicosatetraenoic myeloperoxidase (CCL2), 2 (LC spectrometry mass (LxA deuterated (LTB (HEPE), acid (CXCL1), 1 ligand hydroxyeicosapentaenoic ischemia (HETE), (HDoHE), acid hydroxyeicosatetraenoic acid hydroxydocosahexaenoic (FPR), receptor epoxyeic (EPA), acid eicosapentaenoic dihydroxy cyclooxygenase (COX), (BSA), albumin serum bovine (AA), acid arachidonic (ACN), Acetonitrile 5 2 N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal ), , rsalni E prostaglandin ),

limit of detection (LOD), limit of quantification (LOQ), quantification of limit (LOD), detection of limit Accepted (SMA), artery mesenteric superior (RvD2), D2 resolvin (RvD1), D1 resolvin Article 2

4 (TXB

- d5),

a crmtgah ( chromatography gas leukotriene B leukotriene 910-923. ,DOI :10.1111/bph.12957 - eicosatetraenoic acid (DiHETE), docosahexaenoic acid (DHA), (DHA), acid docosahexaenoic (DiHETE), acid eicosatetraenoic 2 ioi B lipoxin Comment citer cedocument: - ), ultra chromatography high), ( pressure liquid MS/MS), 3

oxo (PGE - 1 ETE),

(PGA 4 3

, 6 ), 7(S)

(LxB (LTB

- keto 1 phosphate osatrienoic acid (EET), formic acid (FA), acid formic (EET), acid osatrienoic maresin (7 maresin ), 8 ), 4 4 ), ), leukotriene B leukotriene ), - - lipoxygenase rsalni F prostaglandin iso prostaglandin A prostaglandin iso GC), GC), neonatal necrotizing enterocolitis (NEC), (NEC), enterocolitis necrotizing neonatal 3 - - ufrd saline buffered rpruin (I/R), /reperfusion MaR1), methanol ( methanol MaR1), aks aacd at ouin (HBSS), solution salt balanced Hank’s

( 4 LOX),

deuterated (LTB deuterated 1 

(6kPGF 2

iud chromatography/tandem liquid lipoxin A lipoxin - ( iso PGA iso PBS UHPLC)

MeOH hmkn (C chemokine 1  ), polyunsaturated fatty fatty polyunsaturated ), , 10(S),17(S) ),

4 4 2 - ),

), prostaglandin E prostaglandin ), d4), leukotriene B leukotriene d4), . (LxA

chemokine ligand ligand chemokine

formyl peptide formyl 4 ), lipoxin A lipoxin ), - X - - motif) C protectin 2 5 4

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 49 48 47 46 45 44 43 64 63 62 61 60 59 58 57 56 55 54 53 52 51 50

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, Summary This article isprotectedbycopyright. Allrights reserved. endogenous production be can influenced by diet. I/R intestinal against protection significant Implications: and Conclusions rich di diet control high a fed group I/R diet control to compared I/R, n high a fed Animals animals. operated increased and inflammation and caused (COX) I/R cyclooxygenase intestinal mice, diet-fed control In results: Key PUFA I/R. performed by inintestinal tissues chromatography liquid tandem mass spectrometry. intestinal after evaluated was injury Intestinal PUFA different two and control, approach: Experimental or n n in rich diet a of consequences the determine to was study this of aim The unknown. is injury chronic purpose: and Background - 6 polyunsaturated fatty acids (PUFA) on intestinal I/R6 polyunsaturatedfatty acids (PUFA) onintestinal N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal et

inflammation, but their potential influence on acute intestinal ischemia/reperfusion (I/R) (I/R) ischemia/reperfusion intestinal acute on influence potential their but inflammation, - Accepted antiassociated Article - fed mice. Blockade of Blockade mice. fed

Lipoxin A Lipoxin 910-923. ,DOI :10.1111/bph.12957 - 3 diet did not display a different inflammatory profile following intestinal following profile inflammatory different a display not did diet 3 n Comment citer cedocument: - inflammatory effects.inflammatory - de ad ee o LxA of level and diet 6 Mice were fed 3 different isocaloric diets: a balanced diet used as a as used diet balanced a diets: isocaloric different 3 fed were Mice Long-term intake of dietary fatty acids is known to predispose to to predispose to known is acids fatty dietary of intake Long-term 4 -

fe (LxA lipoxygenase d animals. In contrast, intestinal inflammation was decreased in decreased was inflammation intestinal contrast, In animals. d This study indicates that high n-6, but not n-3, PUFAs leads to to leads PUFAs n-3, not but n-6, high that indicates study This - nihd it, rvdn ete hg n high either providing diets, enriched

formyl 4 ) was significantly and selectively and significantly was ) - peptide receptor peptide

( -

LOX) nue dmg ad eosrts ht LxA4 that demonstrates and damage induced 4

- a icesd post increased was derived - induced - 2 , the LxA the , metabolites, metabolite quantification was was quantification metabolite

damage. damage. 4 increased after ischemia. ischemia. after increased

- receptor, abrogated receptor, ischemia,

o hg n high or 3 oprd o sham to compared

oprd to compared

- intake. 6 n - - 6 3 -

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 71 70 69 68 67 66 65 87 86 85 84 83 82 81 80 79 78 77 76 75 74 73 72

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, Introduction Introduction This article isprotectedbycopyright. Allrights reserved. microcirculation the to due injury I/R to prone t Therefore, treatments. pharmacological established of lack the to especially and diagnosis in difficulty to Eltzschig transplantation bowel small and enterocolitis cardiovascular necrotizing major surgery, shock, septic hernias, occlusion, vessel including diseases of number of thegutI/R injury. to 3/n damage inflammatory with correlated which n in differences quantitative and temporal identified (I/R) reperfusion ischemia intestinal murine of model experimental LC by characterized, we study, previous our In tissues. in metabolites PUFA of number a of presence the simultaneously disease of comprehension better a allows MS/MS) chromatography liquid sensitivity high of techniques new thrombosis, injury atherosclerosis, (I/R) reperfusion ischemia include arrhythmia, These pathologies. inflammatory and cardiovascular pro engaging or response inflammatory n - plustrtd at ai (PUFA) acid fatty polyunsaturated 6 - 6 ratio composition in membranes could be used as therapeutic tool to modify the resistance resistance the modify to tool therapeutic as used be could membranes in composition ratio 6 N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Acute is There

Acceptedal. et Article he need for new new for need he eetrc shma s life a is ischemia mesenteric , 2011; Vollmar 2011; ,

growing evidence, which indicates bioactive lipid mediators derived from n-3 or or n-3 from derived mediators lipid bioactive indicates which evidence, growing (Vollmar 910-923. ,DOI :10.1111/bph.12957

Comment citer cedocument:

SM, h tsu poie f n of profile tissue the MS/MS, et al. et therapeutic strategies is urgent. The intestinal mucosa is particularly is mucosa intestinal The urgent. is strategies therapeutic

et al. et , 2011) ,

ntmcl n pyilgcl hrceitc f h villus the of characteristic physiological and anatomical , 2011) , . The temporary interruption of the blood leads to a lack lack a to leads blood the of interruption temporary The . play a pivotal role in initiating and sustaining the the sustaining and initiating in role pivotal a play . Intestinal ischemia rema ischemia Intestinal . - resolving/anti . However, it However, . - hetnn ptooia eet soitd with associated event pathological threatening (De Caterina, 2011) Caterina, (De 5 - - soitd iii mtblm detecting metaboloma lipidic associated ad n and 3

- - is unclear whether modulating the n the modulating whether unclear is nlmaoy ahas in pathways inflammatory and 6 - adm as spec mass tandem - PF mtblt production, metabolite PUFA 6

n . - (Gobbetti PF mtblts n an in metabolites PUFA 3 of development recent The ins a clinical challenge due due challenge clinical a ins (Cerqueira

et al. et

rmty (LC trometry t al. et , 2013) , 2005; , several . We We . a - -

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 105 104 103 102 101 100 94 93 92 91 90 89 88 99 98 97 96 95

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, of oxygen supply to the tissue (ischemia), which in turn causes cellular dysfunction, protease and proteasedysfunction, causescellular turn in (ischemia),which tissue the to supplyoxygen of This article isprotectedbycopyright. Allrights reserved. I/Ragainst intestinal LxA of mobilization early that indicates study This diet. rich formyl LxA of blockade pharmacological systemic Early diet. balanced a with fed group the fed group the in decreased was inflammation Intestinal metabolites. PUFA intestinal and PUFA hepatic of abundance the changed significantly intervention Dietary injury. I/R intestinal on metabolites al. n in rich diet dietary a to mice exposing a by intervention performed We injury. reperfusion ischemia to resistance intestinal improve might b metabolite PUFA ofby inducedmodification the status re response, organ and leading translocation multiple tobacterial failure tissue consequent syste and damage) the (epithelial/endothelial local and the aggravates flow (reperfusion) blood of restoration the Paradoxically, p opoiae activation hospholipases 21; Kelavkar 2013; , N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal - The etd receptor peptide

Accepted high a with Article

main objective of this study was to identify if the modification of of modification the if identify to was study this of objective main

- 910-923. ,DOI :10.1111/bph.12957 induced damage. induced t al. et n Comment citer cedocument: - - p2, boae te anti the abrogated (Fpr2), 2 6 diet and the level of of level the and diet 6 (Gobbetti , 2006)

n w eautd h iprac o n of importance the evaluated we and

t al. et

21; Otamiri 2012; , 6 LxA - 3, n 3, 4 -

- 6 PUFA or to a balanced diet balanced a to or PUFA 6 after ischemia was increased compared to to compared increased was ischemia after inflammatory effects attained by the the by attained effects inflammatory iosynthesis associated with diet changesdiet with iosynthesis associated

t al. et 4

18; Vollmar 1987; , leads to significant protection protection significant to leads (Cerqueira

- mic inflammatory inflammatory mic ad n and 3

et al.

n t al. et - - oxygenation oxygenation (Ducheix 6/n , 2005) 4 - 6 PUFA PUFA 6 receptor, - 2011) , 3PUFA 3PUFA . n

- et 6 .

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 124 123 122 121 120 119 118 117 116 115 114 113 112 111 110 109 108 107 106 128 127 126 125

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, MATERIALS AND METHODS ANDMATERIALS METHODS This article isprotectedbycopyright. Allrights reserved. 5,6 EET, 14 and HDoHE) 12 and HETE) dihydroxy HEPE), (LTB deuterated 7(S) (RvD2), A A prostaglandin 6 0.9%. (NaCl) chloride sodium / 1%) (DMSO sulfoxide dimethyl in dissolved was Switzerland), N Chemicals with the ARRIVEguidelines accordance in reported were and council, European the of Animals Laboratory of Use and Care Gui with accordance in conducted were experiments animal All study. this in included No. (permit protocol study whole the approved (CEEA US006/CREFE of Committee ethic and Care Animal water. and food to access 20 at room free pathogen specific a in cage) per mice (4 cages ventilated in maintained were Animals France). Isle, Saint Genest (Le Janvier from purchased were weeks-old) (3 mice male BL/6 C57 Animals

- -tert-butoxycarbonyl-L-Phe-D-Leu-L-Phe-D-Leu-L-Phe (Boc2; Bachem, Bubendorf Bubendorf Bachem, (Boc2; -tert-butoxycarbonyl-L-Phe-D-Leu-L-Phe-D-Leu-L-Phe 4 keto

(LxA 24°C and relative humidity (40% humidity relative and –24°C N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal - rsalni F prostaglandin 4 ), lipoxin B lipoxin), -

Accepted Article E, 5 EET, - maresin (7 maresin - EE 8 HETE, 1

- (PGA 4 DH, 14,15 HDoHE, - - 4, 10(S),17(S) d4), ioaereoc cd (5,6 acid eicosatetraenoic 910-923. ,DOI :10.1111/bph.12957 - 4 xecsttani ai (5 acid oxoeicosatetraenoic

(LxB Comment citer cedocument: 1 1 ), 8 ),  - -

MaR1), leukotriene B leukotriene MaR1), EE 5 HETE, (6kPGF 4 - iso prostaglandin A prostaglandin iso ), lipoxin A lipoxin), (Kilkenny - pxecstini aci epoxyeicosatrienoic - 1

EE 5 HETE,  - , hobxn B thromboxane ), rtci (D) 1 (PDx), protectin

4 et al. –

deuterated(LxA 70%) with a 12 hours light/dark cycle and given fre given and cycle light/dark hours 12 a with 70%) , 2010) - HETE - iEE, 15 DiHETE), 7 4 2

(8 (LTB -

- oxo . - iso PGA iso MP/01/64/09/12

8 17 d8, 4 - ), leukotriene B leukotriene ), T) ee ucae fo Cayman from purchased were ETE) 2 8 4

- (14,15 d - d5), resolvin D1 (RvD1), resolvin D2 D2 resolvin (RvD1), D1 resolvin d5), - (TXB yrxecspnani ai (18 acid hydroxyeicosapentaenoic hydroxy 2 - yrxecsttani ai (15 acid hydroxyeicosatetraenoic ), prostaglandin E prostaglandin ), 2 , rsalni E prostaglandin ), - E) n 11,12 and EET) . n oa 10 ie were mice 160 total In ). - ooaeani ai (17 acid docosahexaenoic 5

(LTB

5 3 ), leukotriene B leukotriene ),

(PGE - 2 E, 8,9 EET, 3 de for the for de

), lipoxin ), (PGE - 122) 122) 2 ), e - - 4 - -

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 148 147 146 145 144 143 142 141 140 139 138 137 136 135 134 133 132 131 130 129 151 150 149

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, Chemicals (Ann Arbor, MI, USA). MI, Arbor, (Ann Chemicals This article isprotectedbycopyright. Allrights reserved. after hours 5 sacrificed were groups Sham reperfusion. I/R of Mice period. ischemic the after immediately removed gently was clip the minutes 50 After 0.9%). (NaCl solution saline with soak gauze with protected incision the and abdomen mice in replaced were SMA and bowel Small ischemia. cause to France) Ulis, u occluded temporarily was (SMA) artery mesenteric superior the and left the to retracted was bowel small the laparotomy, abdominal reaction. animal's of the check adequacy to testing The pinch surgery. toe a of with monitored period was anesthesia all the during anaesthesia under kept mg/kg. were 50 Animals pentobarbital sodium of injection intraperitoneal an by anaesthetized were Mice differentconditions havesurgical subjectanimals/day been the diet protocol(24 to 5days). for an 2 day surgery each For group). per (n=10 (I/R) ischemia/reperfusion (iiii) and I/R) (sham ischemia/reperfusion sham (iii) ischemia (ii) ischemia, sham (i) groups: experimental “mini 5 of number a into experiment the split to used been has design block complete randomized a weeks, 9 After determined. gain weight the and diet the during week a once weighed were Mice PUF different two against control, n both providing diet balanced A weeks. 9 for (w/w) fat diets isocaloric 3 of one to wereassignedmicerandomly twenty andhundred One and surgical design Study procedure Receptors and Channels - N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal e Accepted different 4 in split was group diet each surgery the of days Article The xperiments”. - operated animals, in which abdominal laparotomy and artery isolation were isolation artery and laparotomy abdominal which in animals, operated 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument: allowing reperfusion. Mice of the ischemia groups were sacrificed sacrificed were groups ischemia the of Mice reperfusion. allowing (Alexander A

Molecular target nomenclature conformed to BJP's Guide to Guide BJP's to conformed nomenclature target Molecular - et al. enriched diets, providing either high n high either providing diets, enriched , 2011) ig mcoaclr lp Hrad paau, Les Apparatus, (Harvard clip microvascular a sing 8 .

- ad n and 3 - 6 fatty acid was used as a a as used was acid fatty 6

- 3 or high n high or 3 containing 5% containing5% imals of the 3 the of imals

Following Following - 6 intake. intake. 6

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 171 170 169 168 167 166 165 164 163 162 161 160 159 158 157 156 155 154 153 152 174 173 172

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, performed without occlusion of the vessel, served as controls and were sacrificed just after the the after just sacrificed were and controls as served vessel, the of occlusion without performed This article isprotectedbycopyright. Allrights reserved. animal cage oxidative degradation of toavoid lipids. at vacuum under stored were Pellets 2. composition acid France). (Quimper, Polaris from (n diet enriched acid fatty (n oil seed grape (Ctrl), diet Control the as designed for (50/50) oils were colza and and seed grape were (w/w) preparation diets experimental fat for used Oils 5% Josas). contained and isocaloric were diets three The described previously Fatty diets composition of by followed i.p. pentobarbital cervical dislocation of overdose lethal a by collection sample of purpose the for intravenous by treated µg/kg). (500 Boc2 with vein tail the of injection were group per mice Ten described. previously as I/R and ischemia sub different 4 in randomized were n animals high providing diet the with fed were mice Eighty ischemia reperfusion. intestinal during activation endogenous Fpr2 of was effect experiment the independent determine An to performed (32°C). plate warming a on placed were mice procedure two procedure, surgical the After time. I/R) (sham I/R or ischemia) (sham ischemic corresponding - ae strs lsd h mdie niin f h admnl al During wall. abdominal the of incision midline the closed sutures layer N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article expressed in percentage of total polyunsaturated fatty acids is provided in table in provided is acids fatty polyunsaturated total of percentage in expressed

910-923. ,DOI :10.1111/bph.12957 (Ducheix Comment citer cedocument: - 3). The EPA enriched fish oil that was used in this study was obtained obtained was study this in used was that oil fish enriched EPA The 3).

t al. et The full composition of the diet is given in table 1. table in given is diet the of composition full The

2013) , - 6), and colza oil oil colza and 6), - 0C Te it wr cagd wc a ek n each in week a twice changed were diets The 20°C. Ples ee rprd y PE IR, oy en Jouy (INRA, UPAE by prepared were Pellets . - rus f 0 nml: hm shma sa I/R, sham ischemia, sham animals: 20 of groups 9 ls were euthanized at the end of the study the of end the at euthanized were Animals

/ fish oils (80/20) for a long chain n chain long a for (80/20) oils fish / - i 6 tk. fe 9 weeks, 9 After ntake.

the surgical surgical the The fatty fatty The - 3 Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 194 193 192 191 190 189 188 187 186 185 184 183 182 181 180 179 178 177 176 175 197 196 195

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, DAPI (Invitrogen). As control procedures,reac control DAPIAs (Invitrogen). Prolong with mounted were Slides hours. 2 for France) Pontoise, Cergy Invitrogen, washes anti or France) Lille, anti with along Germany) Heidelberg, Cruz, Fpr2(1/100, Santa againstantibody primarydirected with the overnight at 4°C a on (PBS), Triton X 0.5% mounted and cryostat, saline a buffered in phosphate sections µm in 5 washed into were cut Slides cryoprotected, Scientific). were Fisher Intestines (Thermo slide Superfrost Immunohistochemistry CytometricBeadBiosciences) Array (BD asdescribed previously (CXCL1) 1 ligand motif) an as used was and tissue ileum the in assessed was activity (MPO) Myeloperoxidase (Motta previouslygranulocytedescribed asof infiltration index possible scoremaximum (Cattaruzza of11 as previously described a with summed were feature each for scores The depletion. cell goblet of absence or presence and abscesses, crypt of absence or presence thickening, muscle of of degree extent infiltration, and cellular presence architecture, mucosal normal of destruction of extent graded: were features following the which in system scoringquantitative semi a on based was and investigator the to blinded performed was score damage histological Microscopic eosin. and hematoxylin This article isprotectedbycopyright. Allrights reserved. Assessment inflammation of N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal , Specimens of the ileum were embedded in paraffin. Sections (5 μm) were stained with stained were μm) (5 Sections paraffin. in embedded were ileum the of Specimens

Accepted(Alexa antibody secondary appropriate with incubated were slides Article 910-923. ,DOI :10.1111/bph.12957 - - 100, and 1% bovine serum albumin (BSA) solution (Sigma) and incubat (Sigma)albumin (BSA) solution serum bovine and 1% 100, Ly Comment citer cedocument:

- - and B2 140 Bio (1/400, 6B.2 cytokeratin 18 (1/500, Santa Cruz), anti Cruz), Santa (1/500, 18 cytokeratin chemokine ligand 2 (CCL2) 2 ligand chemokine

- a AD eoe GmbH, Serotec AbD Rad tions were performed with omission of primaryof omission or werewith performed tions 10

were measured in intestinal tissue by tissue intestinal in measured were et al. et (Motta

- et al. CD45 (1/500, R&D systems, R&D (1/500, CD45 , 2012). C 2012). ,

, 2006).

omr Fac) After France). Colmar, et al. et

Fluor , 2012) hemokine (C hemokine

488 and 555; and 488 ® .

containing containing

- X - ed C Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 217 216 215 214 213 212 211 210 209 208 207 206 205 204 203 202 201 200 199 198 220 219 218

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, a aaye b gas by analyzed was microl One acetate. ethyl µL 50 in dissolved and dryness to evaporated was phase organic The w (3:1). hexane/water with heptane 100°C, extracted were at FAMEs of min the and 150 dryness, 1ml for to v/v) evaporated (MeOH,1:20, in methanol in extract BF3 of lipid mL 1 The with transmethylated standard. internal an as glyceryl of presence triheptadecanoate the in extracted lipids were tissue v/v), of mg (2:1, 2 of EGTA equivalent an mM to corresponding methanol/5 in samples tissue of homogenization following (Zadravec previously described as analyzed were acids fatty Hepatic intestinal Hepatic and fatty acid profiling gene of toexpression the was normalized (QT00173551), UK Ltd, I Green SYBR a using 480 LightCycler a with performed was Amplification ( III SuperScript and oligonucleotides from RNA Total UK Real-time PCR analysis quantification of fluorescence using determined software was Metamorph (Molecular Devices). Göttingen, Inc., MicroImaging Zeiss and stained (Carl of Images Germany). configuration inverted the in objective 63X This article isprotectedbycopyright. Allrights reserved. with microscopes confocal LSM-710 Zeiss using acquired were Images antibodies. secondary ) N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal n 1g f h ttl N ws hn reverse then was RNA total the of 1µg and

Accepted primer: mouse following the of one and ) Article alox15 910-923. ,DOI :10.1111/bph.12957 -

iud hoaorpy n 59 Hewlett 5890 a on chromatography liquid Comment citer cedocument: (QT00111034) or (QT00111034)

small intestine intestine small control slides were collected and processed identically. processed and collected were slides control

Invitrogen), Invitrogen), a ioae wt TIo reagent TRIzol with isolated was Hprt alox5

gene, using the 11

. (QT00258622)

codn o h auatrrs instructions. manufacturer's the to according fpr2 - transcribed with random hexamer hexamer random with transcribed

ΔΔ ( QT00171514, Relative expression of the target the of expression Relative C t

method method - akr sse (Hewkett system Packard t al. et

(Boue (Invitrogen

Master kit kit Master Qiagen 21) Briefly, 2010). ,

iter of FAME FAME of iter et al. ),

, 2011) alox12b ( Paisley, Paisley, Qiagen Qiagen

The The as

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 240 239 238 237 236 235 234 233 232 231 230 229 228 227 226 225 224 223 222 221 243 242 241

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, 5,6 12 HETE, PGE of quantification the performed we technique this By chromatography/tandemmeasurements Liquid mass (LC-MS/MS) spectrometry at stored and MeOH with chromatography/tandemmass spectrometry measurements. dissolved were samples under and evaporated was Solvent nitrogen mL). (2 MeOH with eluted were mediators lipid and mL) 2 H with washed was plate the loading, complete After mL/min. 0.1 of rate flow France). Hoerd, Nagel, (Macherey collected, were Supernatants 4°C. at H in min mL 2 to completed 15 for rpm 3000 at centrifuged were Samples quant protein for withdrawn is 10µL s), (LxA mixture standard internal of µL 5 and Invitrogen) (HBSS, solution salt balanced Hank's of µL 500 in biomedical) (MP Instrument FastPrep®-24 a with crushed were intestines small Mouse extraction Intestinal lipid the detectorand respectively. 245°C, at were 225 and injector The bar). (0.5 hydrogen was gas carrier the and 2°C/min, of rate a at 220°C to 110 0.25 i.d., mm This article isprotectedbycopyright. Allrights reserved. 0.32 X mm (30 column capillary fused-silica Famewax a using USA) CA, Alto, Palo Packard, - EET and 5 and EET 3 4 LxA , N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal - d5, LTB d5, -

Accepted8 HETE, Article 4 LxB ,  - m film thickness; Restek, Belfast, UK). Oven temperatur Oven UK). Belfast, Restek, thickness; film m 4 oxo - d4 and 5 and d4 4 - RD, v2 7 RvD2, RvD1, , - ETE ETE HETE, 5 HETE, 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument: 2 in mouse intestinal tissue intestinal mouse in O and submitted to solid to submitted and O - HETE - HETE, 17 HETE, - d8 at 400 ng/mL in MeOH). After 2 crush cycles (6.5 m/s, 30 m/s, (6.5 cycles crush 2 After MeOH). in ng/mL 400 at d8

Briefly, after plate conditioning, the sample was loaded at at loaded was sample the conditioning, plate after Briefly, - a1 LTB MaR1, - HDoHE, 14 HDoHE, ification and 300 µL of cold methanol was added. added. was methanol cold of µL 300 and ification 12

- 4 (Le Faouder (Le phase extraction using HRX using extraction phase LTB , - HDoHE, 14,15 HDoHE, 6kPGF 5

Px 18 PDx, , 1  , TXB ,

et al. et - 2 EET, 11,12 EET, , 2013) , , PGE , - EE 5,6 HEPE, e was programmed from from programmed was e

2 - . To simultaneously To . , PGA , 0 C o liquid for °C 80 2 O/MeOH (90:10, O/MeOH - - 50 mg 96 mg 50 EET, 8,9 EET, - iEE 15 DiHETE, 1 , 8 , - isoPGA - EET, - well well 2 - , Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 263 262 261 260 259 258 257 256 255 254 253 252 251 250 249 248 247 246 245 244 266 265 264

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, algorithmagglomeration criterion. as z (www.r R with obtained were map heat and clustering Hierarchical analysis. the during detected), peak (no interference no showed injection their and evaluated, were samples blank Importantly, considered. not were LOQ the noi to signal a to leading concentration lowest the to corresponded LOQ and 3 over noise to signal a to leading concentration lowest the to corresponded LOD The (S/N). ratio noise to signal using compounds 27 the for determined were (LOQ) quantification of limit the and (LOD) detection of limit The compound. each for applied was 1/X of factor weight a with regression linear A ng/mL. 500 to ng/mL 0.95 from ranging concentration at solutions analysis Quantitative Hunter Techn Mass (Agilent software using done were analysis quantitative detection, and Peak integration conditions. optimized with mode (MRM) Monitoring Reaction Multiple in 0.35 was rate flow The at set was autosampler ThemL/min. min. 12 at B 0% and min 10.5 at B 100% min, 9.5 at B 100% min, 8.5 at B 85% min, 0 at B 0% follows: as was gradient linear The (B). v/v) (100:0.1, FA ACN, and (CAN) acetonitrile water, of consisted phases SB ZorBAX using electro (Ag MS quadrupole triple 6460 Agilent to coupled Infinity) on performed This article isprotectedbycopyright. Allrights reserved. was analysis LC-MS/MS standards, internal deuterated 3 and interest of lipids 27 separate - crs n cutrd ae o 1 on based clustered and scores N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal - Accepted Article Reverse mode. negative in operating ionization spray ultra high performance high ultra - 1 clm (gln Tcnlge) ih gain elution. gradient a with Technologies) (Agilent column C18 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument: lge) Fr ah tnad clbain uvs ee ul uig 10 using built were curves calibration standard, each For ologies). - projec

Pearson correlation coefficient as distance and the Ward Ward the and distance as coefficient correlation Pearson 5°C and the injection volume was 5 µL. 5 was volume injection the and 5°C

liquid chromatography system (UHPLC, Agilent LC1290 Agilent (UHPLC, system chromatography liquid t.org). PUFA metabolite quantities were transformed to to transformed were quantities metabolite PUFA t.org). 13

omc cd F) 7:501vvv () and (A) (75:25:0.1;v/v/v) (FA) acid formic ilent Technologies) equipped with equipped Technologies) ilent - phase UHPLC was performed performed was UHPLC phase se over 10. over se

Data were acquired acquired were Data

All values under under values All The mobile mobile The -

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 273 272 271 270 269 268 267

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, at P≤0.05. 1 measures 2 unpaired Between distributed. normally were data All software. 5 Prism GraphPad using performed wereAnalyses SEM. ± mean as presented are Data This article isprotectedbycopyright. Allrights reserved. Statistical analysis N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal

Accepted Article- - ald t tailed way ANOVA followed b followed ANOVA way - et Mlil cmaios ihn rus / wr promd y repeated by performed were I/R groups within comparisons Multiple test. 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument:

y Tukey’s procedure. Statistical significance was accepted was significance Statistical procedure. Tukey’s y - ru cmaios ee efre b Student’s by performed were comparisons group 14

- Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 293 292 291 290 289 288 287 286 285 284 283 282 281 280 279 278 277 276 275 274 296 295 294

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, mucosa and red blood cells were found in the lumen. Villi lumen. the in found were cells blood red and mucosa of fragments damage: microscopic by characterized was This inflammation. In ofischemia diet, 50minutes animalscontrol fed followedofreperfusion the by 5hours acute in resulted Long-term intestinal diet n-6 prevents weekly foundtobe regardless and assessed was similar it ofdiet intervention dietary During acid. fatty polyunsaturated intestinal of abundance (AA) acid arachidonic n C20:2 group this in n C22:6 (DHA, n C18:3 n on fed mice of intestine the In 3). (Table group diet control n C20:2 3) (Table gut the of same the was content an 3 increased acid composition, lipid (DHA) docosahexaenoic and (AA) arachidonic the between ratio the group, control the to Compared on intervention dietary the of effect the investigate to order In hepatic and intestinal theEffect dietary of abundance on intervention acids. fatty of This article isprotectedbycopyright. Allrights reserved. Results imals receiving the n the receiving imals N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal : : - - , rcioi ai (A C04 n C20:4 (AA, acid arachidonic 6, Accepted (EP acid eicosapentaenoic 3), Article - 3) - od n h lvr f nml rciig h n the receiving animals of liver the in fold were significantly increased compared to the control diet. On the other hand, hand, other the On diet. control the to compared increased significantly were . In the intestine of mice fed with n with fed mice of intestine the In . 910-923. ,DOI :10.1111/bph.12957 the dietary intervention had a major impact on the relative FA composition composition FA relative the on impact major a had intervention dietary the

Comment citer cedocument: at ais F) ee rfld y a crmtgah ( chromatography gas by profiled were (FA) acids fatty - , 2: n C20:3 6, n dcshxeoc cd (DHA) acid docosahexaenoic and 3 diet had a significantly decreased ratio. Even if the if Even ratio. decreased significantly a had diet 3 -

6 and AA were significantly decreased. decreased. significantly were AA and 6 , 2: n C20:5 A, ischemia injury reperfusion - ) ee infcnl icesd oprd o the to compared increased significantly were 6) 15

- ) C25 n C22:5 3), - 6 di 6

were completely denuded or severely or denuded completely were mirrored the changes in the relative relative the in changes the mirrored et, linoleic acid (LA, C18:2 n C18:2 (LA, acid linoleic et,

- (Supplementaryfigure 1) - 6 diet (Figure 1). Conversely, Conversely, 1). (Figure diet 6 3 diet, diet, 3 - 3 and docosahexaenoic acid acid docosahexaenoic and 3

- ioei ai (ALA, acid linolenic The ratio between between ratio The total intestinal FA intestinal total weight gain was was gain weight C analysis GC)

- 6), 6), . Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 316 315 314 313 312 311 310 309 308 307 306 305 304 303 302 301 300 299 298 297 319 318 317

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, shmarpruin n rus f ie umte t cnrl it Fgr 3) 8/15/12 3A). following (Figure diet maximal control to was submitted mice eicosanoids of groups of in ischemia/reperfusion quantity the that was cluster this of characteristic (LOX) ( lipoxygenase cyclooxygenase by metabolism acid arachidonic of product by composed was cluster first The clusters. different 4 formed metabolites PUFA PUFA of n Ctrl, average fed mice of group the different the of proteins) represented of (pg/mg quantity metabolite columns The 3A). (Figure differences main the reveal quanti not were and eicosanoid the 11,12 influence might acids fatty dietary the much how questioned We damage. induced I/R influence to likely profiles docosanoid reperfusion by ischemia followed or ischemia after metabolites n-3 and n-6 intestinal of production the on diet of Effects inflammatoryconsequencesI/R of damaging injury. long a of administration the that n high a with fed group IR the in decreased significantly were expression intestinal CXCL1/CCL2 ischemia in unchanged were inflammatorymarkers I/R in increased markedly (C chemokine This article isprotectedbycopyright. Allrights reserved. architecture crypt of loss with together damaged, - 6 diet (Figure 2 A 2 (Figure diet 6 - N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal EET, 8,9 EET, - Accepted Article dam microscopic the Conversely, reperfusion. - X - fiable in intestinal tissue. intestinal in fiable EET and 5,6 and EET - C motif) ligand 1 (CXCL1)/ 1 ligand motif) C - 910-923. ,DOI :10.1111/bph.12957 E), compared to control diet group. diet control to compared E), - eie mtblts (LTB metabolites derived Comment citer cedocument: versus - EET were not detectable and PGA and detectable not were EET - term

sham operation in control diet animals diet control in operation sham

n - 6 diet, but not of n of not but diet, 6

PUFA metabolites hierarchical clustering was used to used was clustering hierarchical metabolites PUFA animals receiving a high n high a receiving animals

chemokine ligand 2 (CCL2) 2 ligand chemokine 16

4 LxB ,

Meoeoiae MO atvt and activity (MPO) Myeloperoxidase . COX) Taken together these data demonstrate demonstrate data these together Taken 4 - 5/8/15/12 , 3 diet, protects the gut from the pro the from gut the protects diet, 3

RvD1, RvD2, 7 RvD2, RvD1, - age score, MPO activity and and activity MPO score, age

(PGE 1 , 8 , - - 3 diet and then subjected to to subjected then and diet 3 isoPGA 2 , - EE. h prin The HETE). 6kPGF

(Figure 2 A 2 (Figure

expressi - MaR1, 14,15 MaR1, 2

and 5,6 and 1 α TXB , - ons 3 or n or 3 - E). These These E). - were also were DiHETE DiHETE - - 2 HETE, HETE, 6 diet. diet. 6 ) and and ) - EET, EET, cipal - Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 339 338 337 336 335 334 333 332 331 330 329 328 327 326 325 324 323 322 321 320 342 341 340

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, ic te ise ees f LxA of levels tissue the Since Fpr2 mouse small in isexpressed intestine anti this the ischemia, after Interestingly, expression. lipoxygenase of increase an by ischemia/reperfusion or ischemia after tissue colonic in quantity metabolites PUFA the change to able is supplementation diet that ( ischemia after diet control the receiving mice in increase significant a quantified 3B) (Figure ischemia and diet control to submitted mice LxA ischemia, after diet, significantly our focused we injury, reperfusion ischemia on diet n-6 of effect protective the to Due LxA on attention n to submitted ischemia/reperfusion (Figure 3A). animals in increased were las The diet. the of independently ischemia 15dPGj ischemia sham n to to submitted mice of group in compared maximal ischemia to onl represented was submitted cluster second The 2). Figure group(Supplementary diet control in increased 5 2). Figure (Supplementary I/R sham to compared I/R to submitted This article isprotectedbycopyright. Allrights reserved. PGE 2 , N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal 6kPGF 2 -

inflammatory theprotection in offered diet. lipid by this Accepted Articleby metabolites two these of decrease a by characterized cluster third the forming were increased compared to sham after ischemia (Figure 3B). In mice submitted to n to submitted mice In 3B). (Figure ischemia after sham to compared increased 1 increased LxA increased α , TXB , 4 . In mice receiving the control diet, the anti the diet, control the receiving mice In . 910-923. ,DOI :10.1111/bph.12957 2 , LTB , Comment citer cedocument: 4

was significantly increased compared to compared increased significantly was 4 4

in n in and LxB and

6 diet 6 lx, alox12 alox5, 4 were modified in some of the groups studied, we also also we studied, groups the of some in modified were 4

were significantly increased only increased significantly were vs. - 6 diet and to ischemia (Figure 3 A). 5oxo A). 3 (Figure ischemia to and diet 6 t cluster regrouped all n all regrouped cluster t

control diet suggests a possible protective role for role protective possible a suggests diet control 17 Supplementary Figure 3). 3). Figure Supplementary

- and 3 diet, independently of ischemia or or ischemia of independently diet, 3 alox15 . Moreover, in this group of mice we we mice of group this in Moreover, . - inflammatory mediator LxA mediator inflammatory

RA xrsin oprd to compared expression mRNA

y by LxA by y - sham and also compared to compared also and sham 3 PUFA metabolites. They They metabolites. PUFA 3 - EE a significantly was HETE These findings suggest suggest findings These

in control diet group diet control in 4 ; its quantity was was quantity its ; - ETE and and ETE 4

was - 6 Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 362 361 360 359 358 357 356 355 354 353 352 351 350 349 348 347 346 345 344 343 365 364 363

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, boae as te niiin of inhibition the also abrogated CCL2 and CXCL1 n by activity, induced MPO expression score, damage microscopic of inhibition the abolished n high LxA n theby obtained effects anti-inflammatory the abrogated Fpr2 of blockade Pharmacological control procedures expressin granulocytes of infiltration the of consequence of overexpression the groups, two these in Thus, 4B). Figure (Supplementary n and control in ischemia/reperfusion Ly 2B), (Figure activity MPO with correlation In diets. or groups different the between fluorescence 4A) Fpr2 Figure (Supplementary of observed difference was intensity no types, cell hematopoieti different nucleated those In of 4B). 44% (Figure on respectively and neutrophils cells, epithelial on expressed Ly 18, cytokeratin murine of mucosa in expressed was Fpr2 that observed we immunohistochemistry, By 4A). (Figure sham to of expression the I/R, control in I/R after modify This article isprotectedbycopyright. Allrights reserved. LxA the of expression the investigated - 6 rich diet. N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal - 6 diet, administration of the Fpr pan antagonist, Boc2 (500 µg/kg before ischemia i.v.) i.v.) ischemia before µg/kg (500 Boc2 antagonist, pan Fpr the of administration diet, 6 fpr2

Accepted4 Article is an effector of endogenous anti-inflammation acting anti-inflammation endogenous of effector an is

expression compared to sham (Figure 4A). We observed an increase of of increase an observed We 4A). (Figure sham to compared expression

are shown insupplementaryare shown figure4C. and n and -

B2 n prily ih CD45 with partially and 6B.2 910-923. ,DOI :10.1111/bph.12957 ml itsie Fgr 4) Fpr2 4B). (Figure intestine small fpr2 Comment citer cedocument: - - 3 diet groups compared to corresponding sham (Figure 4A). Following 4A). (Figure sham corresponding to compared groups diet 3 de floig I/R following diet 6

mRNA was not different in mice that received the n the received that mice in different not was mRNA 15

- EE PGE HETE, 4 eetr p2 n h sm tsus Icei dd not did Ischemia tissues. same the in Fpr2 receptor - de gop cmae t IR n I/R to compared groups diet 3 18 Fgr 5) Amnsrto o Fr antagonist Fpr of Administration 5A). (Figure

2 - , B2 xrsin wa expression 6B.2 - immunoreactivity meaning that Fpr2 was was Fpr2 that meaning immunoreactivity 6kPGF

Fr. itrs f immunochemistry of Pictures Fpr2. g - muoeciiy co immunoreactivity 1 α

n TXB and via Fpr2. In mice receiving a a receiving mice In Fpr2. icesd following increased s 2

nue b te n the by induced - fpr2 6 diet, compared diet, 6 - - oaie with localized de group diet 6

could be the be could fpr2

c cells, cells, c mRNA mRNA the the - 6 Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 370 369 368 367 366

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article - - induced protectionI/Rinduced after isindeedFPR2 activation. mediated by 6 diet group diet 6 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument: Administration of Boc2 had no effect on the increase of LxA4 induced LxA4 induced of increase effectonthe Boc2 had ofno Administration (Figure 5C). These findings show that the blockade of the LxA the of blockade the that show findings These 5C). (Figure - inflammatory effects obtained by the the by obtained effects inflammatory

n - 6

rich diet and that n that and diet rich

- 6 4

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 390 389 388 387 386 385 384 383 382 381 380 379 378 377 376 375 374 373 372 371 393 392 391

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, Simultaneously, potent anti potent Simultaneously, (PGE COX by metabolism inc an to led reperfusion by followed ischemia and ischemia that observed we conditions, experimental (Dufton bu lipids, bioactive inflammatory pro of precursor a as only not considered now is AA and studies, of number a by developed response inflammatory resolving in play in dietaryfatty acid. protectivepathwayendogenous n LxA the that demonstrated acid fatty enriched 6 LxA of consequently and expression ofincrease an with associated was I/Rinjury against Protection compar expression, chemokine and recruitment n I/R, intestinal to subjected mice in Specifically, settings. these in determined as damage intestinal on effect I/R against protection significant afforded n and This article isprotectedbycopyright. Allrights reserved. Discussion - 6 PUFA- 6 - N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal 3 PUFA intake, on injury associated with intestinal I/R. Our data indicate that n that indicate data Our I/R. intestinal with associated injury on intake, PUFA 3 es o pro of rease u rsls ute cnimd the confirmed further results Our n- different on based intervention dietary of effect the evaluated we study present the In

Acceptedal. et Article enriched diet. diet. enriched , 2010; Morris 2010; , - 6 PUFA supplementation in diet decreased mucosal damage, granulocyte granulocyte damage, mucosal decreased diet in supplementation PUFA 6

910-923. ,DOI :10.1111/bph.12957 - nlmaoy iatv lpd o ptwy akr eie fo AA from derived marker pathway or lipids bioactive inflammatory - rae gop oprd o oto diet control to compared group treated Comment citer cedocument: Collectively, these data make us propose that early during ischemia, ischemia, during early that propose us make data these Collectively, 4 -

inflammatory mediators derived from the AA metabolism, LxA metabolism, AA the from derived mediators inflammatory receptor Fpr2 was involved in the protective effects associated with associated effects protective the in involved was Fpr2 receptor

2 et al. et 6kPGF , mediated by mediated t also of potent anti potent of also t , 2006; Schwab 2006; ,

production following ischemia in intestinal tissues from n from tissues intestinal in ischemia following production 1  in vivo in

- seta rl th role essential TxB induced tissue injuries tissue induced LxA 20 2 ad O (5 LOX and ) (Serhan 4

d o nml fd cnrl aacd diet. balanced control a fed animals to ed

and its receptor could be triggered by changes triggeredby receptorbe could its and

et al. et - nlmaoymdaosnmd Lipoxins named mediators inflammatory

, 2006; Wallace, 2006) Wallace, 2006; , et al. et t AA at , 1988) , - alox5, alox12 alox5, e group. fed - - 8 , , while n while , derived lipid mediators could could mediators lipid derived - . This notion has been well been has notion This . 12 ,

- - 15 , 3 PUFA diet had no had diet PUFA 3 n vivo In

and - . Indeed, in our our in Indeed, . EE LTB HETE, alox15

experiments experiments - 6 PUFA 6

mRNA 4

and an an 4 ). 6 - - Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 413 412 411 410 409 408 407 406 405 404 403 402 401 400 399 398 397 396 395 394 416 415 414

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, isnhszd y oprto bten 15 between cooperation by biosynthesized I/R in analogues (Bannenberg lipoxin stable or lipoxins delivered pharmacologically for role protective Maderna infect of phagocytosis macrophage migration leukocyte halt to able 2009) 2005) anti endogenous potent a as described synthesis transcellular by principally produced lipid lived suggestsroleLxB afor minor conditions. ischemic LxA that notion the supports and data current our with agreement LxA of neutrophil/pla by activated mechanism a reported recently have We groups. both in same the were discovered), be to yet is receptor(s) (whose n in higher only increased LxA time, over This article isprotectedbycopyright. Allrights reserved. LxB 4 ee lo nrae. hs to iois r dfeetal epesd n netnl tissues intestinal in expressed differentially are lipoxins two These increased. also were , a G protein G a , Fpr2 receptor, its through Acting . N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Cell-cell interaction is one of the routes by which lipoxins are endogenously endogenously are lipoxins which by routes the of one is interaction Cell-cell LxA 4

at the end of 30 min mesenteric ischemia ischemia mesenteric min 30 of end the at Accepted al. et Article - 4 6 diet group after ischemia compared to control diet group. However, levels LxB4 levels However, group. diet control to compared ischemia after group diet 6

was identified by Serhan and colleagues in 1984 (Serhan 1984 in colleagues and Serhan by identified was et

after 5 hours reperfusion. In our study, increased level of level increased study, our In reperfusion. hours 5 after 2005) , al. 4

en poue fo te shmc eid n LxB and period ischemic the from produced being , 2004;Sun - coupled receptor belonging to the formyl peptide receptor family, LxA4 is LxA4 family, receptor peptide formyl the to belonging receptor coupled 910-923. ,DOI :10.1111/bph.12957 h ef The Comment citer cedocument: I areet ih u rsls svrl tde hv dmntae a demonstrated have studies several results, our with agreement In . et f X4Fr sse i te rtcie fet o n of effects protective the in system LXA4/Fpr2 of fect 4

et al.

in ourexperimental conditions. , 2009;Wu

ious agents and apoptotic neutrophils apoptotic and agents ious (Chiang - inflammatory bioactive lipid mediator mediator lipid bioactive inflammatory (Lavigne -

t al. et n 5 and

et al. 21

telet aggregates to increased circulating levels circulating increased to aggregates telet 20; Perretti 2006; ,

- lipoxygenases (LOX) (LOX) lipoxygenases , 2012) et al. et (Brancaleone , 2002; Perretti 2002; .

(Serhan

t al. et 4

t al. et of metabolite early an is 4 et al. et

t al. et

(his (his LxA et al. et 2009) ,

(Serhan (Godson Ti i in is This 2013). , , 1984) as a short- a as 1984) , 4 oiinl isomer) positional LxA 1984). , , 2002; Ye 2002; ,

was significantly was (Serhan

t al. et and promote promote and t al. et 2008) ,

, 2000; , -

6 diet diet 6 t al. et et al. et 4

is . , , Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 436 435 434 433 432 431 430 429 428 427 426 425 424 423 422 421 420 419 418 417 439 438 437

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, eetri IR i th in I/R; in receptor 2010) injury I/R of pathophysiology the in role important an have might FPR2 particular in humanin LXA of properties resolving mice in Fpr2/3 and humans formyl termed receptor target specific gut) the populate physiologically that cells mast and macrophages, leukocytes, or H(p)ETE 15S of source principal the be could cells epithelial infiltration, granulocyte before ischemia where conditions, experimental our to compared patients, colitis ulcerative (Mangino LxA colitis, mucosa H(p)ETE 15 expressed Serhan 5 leukocyte of action the by Serhan 15S form alcohol 15 by AA of metabolism This article isprotectedbycopyright. Allrights reserved. cells, dendritic and cells epithelial airway in later and granulocytes in described Primarily I a eet td, h ue of use the study, recent a In . N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal

et al. et t al. et (Mangino (Dufton

(McMahon Accepted al. et Article

4 , 2008) , -

15s 2008) , O n ol ute otiuet LxA to contribute further could and LOX noeos rdcin s infcnl rdcd oprd o oto patients control to compared reduced significantly is production endogenous , 2006) , -

et al. et - EE eaoie it LxA into metabolized HETE

hydroxyeicosatetraenoic acid (15S acid hydroxyeicosatetraenoic . t al. et

910-923. ,DOI :10.1111/bph.12957 . intermediates epoxytetraene into transformed further are metabolites These

e absence of Fpr2/3, an aberrant inflammatory response characterized by by characterized response inflammatory aberrant an Fpr2/3, of absence e

t al. et . This decrease was coupled to a lower level of 15 of level lower a to coupled was decrease This . . Increasing2010), evidenc . n h intestine, the In Comment citer cedocument: 2006) , - LOX produces 15S produces LOX 2001) , - 4 (Perretti O yedn LxA yielding LOX ,

u also but Itrsigy i pathological in Interestingly, . Ide, LxA Indeed, .

alox5

fpr2/3 t al. et - tissues from healthy controls healthy from tissues etd receptor peptide

resident h guootci rgltd rti Anxn 1 and A1 Annexin protein regulated glucocorticoid the

and 20; Ye 2002; , - deficient mice highlighted the importance of this this of importance the highlighted mice deficient - alox15 22 hydroperoxyeicosatetraenoic acid or the reduced reduced the or acid hydroperoxyeicosatetraenoic 4

e suggests that members of the FPR family FPR and the of members suggeststhat e tissue cells such as intestinal intestinal as such cells tissue 4 4 by

is synthesized in the human gut by colonic colonic by gut human the in synthesized is n LxB and -

5 HETE) expression and LxA and expression -

O baig cells bearing LOX t al. et - 4 /ioi A 2/lipoxin

generation by the production of 15S of production the by generation 4 2009) , (Levy (Levy

conditions such as ulcerative ulcerative as such conditions

t al. et et al. et 4 . FPR2 transduces the pro pro the transduces FPR2 . (Mangino

receptor (FPR2/ALX) in in (FPR2/ALX) receptor - LOX in the mucosa of mucosa the in LOX , 1993; Serhan, 1989; 1989; Serhan, 1993; ,

, 1993; Serhan, 1989; Serhan, 1993; , 4

( were increased after increased were polymorphonuclear polymorphonuclear . LxA .

et al. et epithelial cells cells epithelial 4 , 2006) ,

acts on a a on acts (Gavins, (Gavins, . In . - - Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 459 458 457 456 455 454 453 452 451 450 449 448 447 446 445 444 443 442 441 440 462 461 460

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, f p2 naeet y LxA by engagement Fpr2 of LxA by modulated interleukin n LxA to exposure cell by inhibited pro the release cells epithelial TNF cell epithelial LxA inflammation. intestinal on acting byalso but cells, immune innate regulating by only not of signals inflammatory attenuation potential in resulting side, basolateral leukocyte cells epithelial vitro lines cellepithelial intestinal Fpr2in identified had that study previous a confirming cells, epithelial intestinal in also expressed was receptor murine monocytes and neutrophils n the in higheramount post of n between association protective the that indicate data Our anti protective the reverted ischemia before antagonist Fpr2 the d LxA of delivery exogenous This article isprotectedbycopyright. Allrights reserved. (Brancaleone infiltration and adhesion neutrophil prolonged - animals eficient 6 - enriched diet was associated with a decrease in CXCL1, the murine equivalent of human human of equivalent murine the CXCL1, in decrease a with associated was diet enriched  studies have identified Fpr2 expression preferentially on the basolateral side of intestinal of side basolateral the on preferentially expression Fpr2 identified have studies N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal (Goh - ischemic damage was due, at least in part, to the interaction between LxA between interaction the to part, in least at due, was damage ischemic - pteil el neato c interaction cell epithelial

Accepted- Article . te pro Other 8.

t al. et s. Upon stimulation with bacteria ( bacteria with stimulation Upon s. (Kucharzik (Brancaleone 20; Gronert 2001; , 4

in epithelial cells epithelial in 910-923. ,DOI :10.1111/bph.12957 - Comment citer cedocument: 6 group) and its receptor, Fpr2. Human FPR2/ALX was first identified onidentified firstwas FPR2/ALX Humanreceptor, Fpr2. its andgroup) 6 - nlmaoy inl, uh s t as such signals, inflammatory

et al. et 4 (

Fiore attenuated I/Rattenuated 4

(Gewirtz - et al. et nlmaoy hmkn interleukin chemokine inflammatory , 2003) , 4

. In accordance with these data, we observed that after I/R the I/R after that observed we data, these with accordance In .

et al. et

t al. et , 2013) , (Gewirtz ould rapidly signal by activating the receptor on the the on receptor the activating by signal rapidly ould . LxA . , 1992; Takano 1992; ,

t al. et 1998) , - mediated inflammation in wild type but not in Fpr2/3 in not typebut wild in inflammation mediated . In mice receiving a high n high a receiving mice In . 4

2002). , produced on the serosal side of the epithelium by epithelium the of side serosal the on produced

23 et al. et Salmonella typhimurium Salmonella

r LPS or , 1998) ,

regarding data conflicting is There et al. et e rncito fco NF factor transcription he (Kure - 6 supplementation and the prevention prevention the and supplementation 6 . Such inhibition is also downstream downstream also is inhibition Such . , 1997) , (Goh -

inflammatory effect of this diet. this of effect inflammatory t al. et t al. et

et al. et . Here, we observed that the that observed we Here, . - : hs respon this 8: 2010) , - 6 diet, administration of administration diet, 6 , 2001) , ) , 2013). Furthermore, Furthermore, 2013). ,

(Gewirtz hmn intestinal human , 4 . Interestingly, .

rmts anti promotes 4

et al. et (produced in (produced e ol be could se -  B can be be can B , 1998) , in vitro in how how in in - ,

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 482 481 480 479 478 477 476 475 474 473 472 471 470 469 468 467 466 465 464 463 486 485 484 483

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, injuries reperfusion ischemia the of prevention the for approach therapeutic viable a as supplementation a circuit anti-inflammatory endogenous the that propose to us prompt data these conclusion In treatment. AA than effective more even be could model, our in metabolite AA important most the be injury intestinal of degree the reduce to shown been have AA with formula the of supplementation model, NEC rat the controls with compared PUFA of amounts higher containing formula lo a found al. et Carlson study, clinical a In pathology. this of outcome the regulate can diet infant of composition and type the that shown been has (NEC) enterocolitis necrotizing neonatal as such pathogenesis, the to coischemia intestinal which in diseases in therapeuticimpactof be coulddescribed here Exploiting scenarios. in intestinal specific work may diets n-6 high of consumption the that indicating properties anti-inflammatory displayed diet enriched n-3 the not but n-6 the Conversely, diet. control or n-3 the to compared “pro any detect status inﬂammatory lowest the have LA of level highest the consuming individuals that shown have studies Several responses. higher with associated not is circulation or diet the in and (AA) acid arachidonic proof production subsequent tissue of accumulation through inﬂammation promotes intake LA high that claim Some diet. healthy a in consumed be should LA) acid, linoleic (i.e. n-6 much This article isprotectedbycopyright. Allrights reserved. tvtd y slcie p2 gns, LxA agonist, Fpr2 selective a by ctivated N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal

Accepted Article - inflamamtory phenotype” of the n the of phenotype” inflamamtory 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument: (Lu inﬂammatory lipid mediators. Human studie Human mediators. inﬂammatory lipid

et al. et Fish, 2008) (Fritsche,

, 2007)

this this . Thus in this pathology, delivery of LxA of delivery pathology, this in Thus . 4 n cud e anse b botn n boosting by harnessed be could , - 24 6 dependent 6 I or xeietl nml td w dd not did we study animal experimental our In .

-6 diet group that underwent a sham operation operation sham a underwent that group diet -6 e icdne f E i ifns fed infants in NEC of incidence wer n vivo in

endogenous anti endogenous or or x vivo ex (Carlson (Berman s indicate that high LAhigh that indicate s

- inflammatory loop inflammatory pro

et al. et et al. et - - 6 PUFA diet diet PUFA 6 inﬂammatory inﬂammatory 4 , 1998) , , shown to to shown , , 2011) , ntributes - based based . In . . It . Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 506 505 504 503 502 501 500 499 498 497 496 495 494 493 492 491 490 489 488 487 510 509 508 507

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, from article inception topublished whole, a as work the of integrity the for responsibility take Cenac Nicolas and Gobbetti Thomas of themanuscript Nicolasconception and Cenac: design; analysis acquisition, and interpretation ofdata; redaction Nathalie Vergnolle: ofthe and conception manuscript design;redaction Mauro and conception design; Perretti: redaction of themanuscript Hervé conceptionGuillou: and design;of interpretation data; manuscript redaction of the Marc Dubourdeau: interpretationofdata; Bertrand-Michel:Justine analysisand interpretation ofdata Boué:Jérôme and analysis ofdata acquisition Pauline leFaouder: andanalysis acquisition redaction ofdata; of themanuscript acquisition Ducheix: andSimon analysis ofdata Thomas acquisition, Gobbetti: analysisof ofdata; themanuscript and redaction interpretation author contributions: of List Midi (program 086867/Z/0 the european (to research NV,ERC council 0001 AgenceNationale andthank Dr.construction analyses. heat-map Pascal for Martin facilityandcore thehistology US006/INSERM, Genetoul, Toulouse.Authors facility, anexplo, INSERMThe UMR1043,Toulouse,facility, authorscore the thank the microscopeanimal core This article isprotectedbycopyright. Allrights reserved. Acknowledgements - - Pyrénées and theEuropean Union. 01), the INSERM (to NV), two grantsINSERM NV),01), the theRegion two (to from Midi N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article

de la Recherche la ANRde (to NV,

910-923. ,DOI :10.1111/bph.12957 8/Z, to TG and MP). Ambiotis SAS is supported is by toTGand SAS Ambiotis MP). fromgrants the8/Z, Region Comment citer cedocument:

redaction of the manuscript of themanuscript redaction - 2012 25 - - 12

StG - BSV1 - 20111109) and - 0030

- This work was supported by the This work 01 and NC, ANR and NC, 01 - Pyrénées (to PLFandPyrénées NC the

Wellcome Trust

- 12 - JSV1 - ), Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 549 548 547 546 545 544 543 542 541 540 539 538 537 536 535 534 533 532 531 530 529 528 527 526 525 524 523 522 521 520 519 518 517 516 515 514 513 512 511 557 556 555 554 553 552 551 550

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, lipox S, Fiore Ryeom SW,Weller SerhanPF, CN (1992). Lipoxin recognition sites. Specific binding labeled of 1391 Eltzschig HK,Eckle (2011). T Ischemia and reperfusion agonists. Dufton N,Perretti (2010).M Therapeuticanti-inflammatory potential formyl-peptideof receptor Hepatol acids deficiency promoteslipogenic gene expression and hepaticsteatosis throughliver the receptor.X S,Ducheix Montagner A, Polizzi A,Lasserre F, Marmugi A,Bertrand-Michel J 364 Caterina De R (2011). fatty n-3 acids incardiovascular disease. potent ligand Chiang N,Serhan CN,Dahlen SE, Drazen JM, Hay DW, Rovati GE review. Cerqueira NF, Hussni CA, Yoshida WB(2005). Pathophysiology of mesenteric ischemia/reperfusion: a intestinal ischemia/reperfusion inrodents. proteinase Cattaruzza F,CenacN, Barocelli ImpicciatoreE, M, Hyun VergnolleN E, enterocolitis ininfants fed a preterm formula with egg phospholipids. Carlson SE, Montalto MB, Ponder DL, Werkman SH, KoronesSB (1998). Lower incidence necrotizingof microcirculation. circuit centered lipoxin on andA4 aspirin- BrancaleoneV, Gobbetti T, Cenac N, leFaouder P, Colom B, Flower RJ dependent adaptiveon T cell responsein mice. J, Boue BlanpiedBrousset C, P, VergnolleN, Dietrich G (2011). Endogenous opioid-mediated analgesia is 150. Berman L, Moss RL (2011). Necrotizing enterocolitis: an update. Pharmacol and novel 1 Bannenberg G, Moussignac RL, GronertK, Devchand PR, Schmidt BA,Guilford WJ Pharmacol AlexanderSP, A,Mathie Peters JA (2011). Guide to Receptors and Channels (GRAC),5th edition. This article isprotectedbycopyright. Allrights reserved. References (25) in A4 human with neutrophils. - 1401. N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal :

Acta Cir Bras

58 2439 Pharmacol Ther - (5) 143 164 Suppl 1:

activated receptor activation2 on motility impairment and tissue damageinduced by Accepted Article5

- - epi : - specificand stereoselectiveactions i

2450. 984 (1) - Blood lipoxin analogs display potent anti :

- 43 992.

20 - 910-923. ,DOI :10.1111/bph.12957 52.

122 (4)

Comment citer cedocument:

127 : -

(4) 324. 336 (2) :

608 - 343. :

175 - J Biol Chem 617.

- 188.

triggered 15

AmJ Pathol

267 J Immunol n vivo. 26 (23) - inflammatory actions after oral administration.

-- - epi from mechanism to translation. : 169

16168 Pharmacol Rev -

186 lipoxin A4 operativein murine (1) : (9) The NewEngland journal ofmedicine

- 177 , etal. 16176. Semin Fetal Neonatal Med :

5078 - 188. , al. et (2006). Thelipoxin receptor ALX: Pediatr Res

, etal. - 5084.

58 (3) (2013). Avasculo-protective , et al.

(2006). Protective effectof

463

,(2004). al. et Lipoxins

44 (2013). Essential fatty - 487. (4) : Nat Med

491

16(3) - 498.

Br J : 17 145-

(11)

Br J : J

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 597 596 595 594 593 592 591 590 589 588 587 586 585 584 583 582 581 580 579 578 577 576 575 574 573 572 571 570 569 568 567 566 565 564 563 562 561 560 559 558 603 602 601 600 599 598

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, C888 mediateLXA4's anti Kucharzik T, Gewirtz AT, Merlin D, Madara JL, Williams(2003). IR Lateral membraneLXA4 receptors reporting:ARRIVE the guidelinesfor reporting animal research. Kilkenny C, BrowneWJ, Cuthill IC,Emerson M, Altman(2010). DG Improving bioscienceresearch radical prostatectomy. can be modulated by the omega Kelavkar UP, Hutzley J, DhirKim R, AllenP, McHugh KG, (2006). K Prostate tumor growth and recurrence alpha receptor thatis regulated by interleukin (IL) Gronert K, Gewirtz A, Madara JL, Serhan CN (1998). Identification ofhuman a enterocytelipoxin A4 mucosa ex vivo. vitro and attenuate TNF triggered 15 J,BairdGoh AW, O'Keane C, Watson RW,Cottell BernasconiD, G Immunol stimulatenonphlogi phagocytosisstic of apoptoticneutrophils by monocyte Godson C, Mitchell S,Harvey K, Petasis NA, Hogg N,Brady HR(2000). Cutting edge: lipoxins rapidly e75581. AcidFatty metabolism signaturein ischemia differs from reperfusion in mouseintestine. Gobbetti T, Le Faouder P, Bertrand J, Dubourdeau M, Barocelli Cenac E, N 152. inhibi Gobbetti T, Cenac N, Motta Rolland JP, C, Martin Andrade-Gordon L, P 101 chemokine secretion from model intestinal epitheliuminhibited is lipoxin by A4 analogs. Gewirtz AT, McCormick B, Neish AS, Petasis NA, GronertK, Serhan CN severity of dextran sodium sulfate analogs attenuate induction intestinal of e Gewirtz AT,Collier-Hyams YoungLS, AN, Kucharzik T, Guilford WJ, Parkinson JF reperfusion injury? Gavins FN (2010). Areformyl peptide receptors novel for targets therapeuticintervention inischaemia- Essent FattyAcids This article isprotectedbycopyright. Allrights reserved. Fritsche KL (2008). Too linoleicmuch acidpromotesinflammation-doesn't it? (9) - - tion reducestion post 896. induced IL N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal :

1860

164 Accepted Article - - epi (4) 1869. : - J Immunol -

lipoxin A(4)antagonize TNF 1663 8 release.8

Trends Pharmacol Sci - inflammatory actions intestinalon epithelium. (3 - 910-923. ,DOI :10.1111/bph.12957 1667. - - Neoplasia 5) ischemic granulocyte recruitment inmouse intestine. - alpha Comment citer cedocument: :

173 167 J Med Exp

- - induced chemokine release and colonocyteapoptosis in human intestinal (5) 175. -

6:omega 8 : -

(2) induced colitis. J Immunol 2772

: 187

112 - 2780.

31(6) (8) - pithelial proinflammatory gene expression and reducethe - 3 ratio indiet: athymic mouse xenograftmodel simulating 124. - - 13 and interferon gamma and inhibits tumornecrosis factor : a

1285 lpha

266 - - 27 stimulated neutrophil - 1294. - 276.

168 PLoS Biol , al. et (10) Am J Physiol Cell Physiol : , al.et

, al. et 5260 (2001). Lipoxin A(4)and aspirin-

8 , al. et enterocyte interactions in (6) - - (1998). Pathogen-induced 5267. derived macrophages. (2012). Serine protease : ProstaglandinsLeukot AmJ Pathol

e1000412. , al.et

(2013). Polyunsaturated

(2002). Lipoxin a4 J Clin Invest PLoS One

180 284 (1) (4) :

141

: 8 J

(9)

- :

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 643 642 641 640 639 638 637 636 635 634 633 632 631 630 629 628 627 626 625 624 623 622 621 620 619 618 617 616 615 614 613 612 611 610 609 608 607 606 605 604 651 650 649 648 647 646 645 644

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, inflammation. Perretti M, D'Acquisto F(2009). Annexin A1 and glucocorticoids as effectors of the resolution of lipoxin A4 recept derived anti - Perretti ChiangM, N, La M, Fierro IM, Marullo S,Getting SJ 28(11) lysophospholipase activity insmall the intestinal mucosa after ischaemia and revasculari Otamiri Franzen T, L, Lindmark D, Tagesson C (1987). Increased phospholipase A2 and decreased 4 bacteria expressing elafin protectagainst infla JP,Motta Bermudez-Humaran LG, Deraison C, Martin RollandL, Rousset C, P 27(12) Morris T, RajakariarStables R, M, Gilroy DW (2006). Notall eicosanoidsbad. are Pharmacol Sci McMahon B, Mitchell S, Brady HR,Godson(2001). C Lipoxins: revelations resolution.on ProstaglandinsOther Lipid Mediat Mangino MJ, Brounts HarmsL, B, HeiseC (2006). Lipoxin biosynthesis in inflammatory bowel disease. Immunol by supernatant from dexamethasone Maderna P, S,Yona Perretti Godson M, (2005). C Modulation phagocytosisof of apoptoticneutrophils Res geneexpression and reducesthe incidence of necrotizing enterocolitis inaneonatal rat model. Lu J,Jilling T, D,Li Caplan (2007).MS J Clin Invest have 15 BD,Levy Romano M, Chapman HA, Reilly JJ, Drazen J, Serhan CN(1993). Human alveolar macrophages metabolites. rapid and concomitant quantification proof Faouder Le BaillifP, V,SpreadburyI, Motta JP, Rousset Chene P, G Cell Immunol coupled receptor mediate fMet Lavigne MC, Murphy PM, Leto TL, Gao JL (2002). The N-formylpeptide receptor (FPR) and a second G(i)- inhibition nuclear of factor lipopolysaccharide This article isprotectedbycopyright. Allrights reserved. Kure I,Nishiumi S,NishitaniTanoue Y, Ishida T,T, Mizuno M (158)

61(4) N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal : : :

1445 158ra144. 609 - lipoxygenase and generate 15(S)

: 174

427 - Accepted Article 92 611. - inflammatory pathways generated with glucocorticoid and aspirin treatment activate the

J Chromatogr B AnalytTechnol Biomed Life Sci (6) 1453. 218

- 22(8) (3) Nat Rev Immunol 432. :

or. 3727 : (1

1572 -

induced inflammation in macrophagesand intestinal epithelial cells through : 2) Nat Med

391 : - 910-923. ,DOI :10.1111/bph.12957

3733. 7 - 1579. - - Comment citer cedocument: 12. 395. - kappaB activation.

(11) - - Leu 9 (1)

79(1 :

1296 : Phe Pol

- 62 treated macrophagesand annexin

- 2) yunsaturated fatty acidsupplementation alters proinflammatory - - 70. - stimulated activation NADPHof oxidase inneutrophils. murine - 1302. hydroxy :

84 -

inflammatory and pro - mmation and restore colon homeostasis. 92. J Pharmacol Exp Ther

- 5,8,11 28

- cis , et al. , et

932C: , al. et - 13- trans- (2002). Endogenous lipid-and peptide-

123 , et al. (2010). Lipoxinreduces A(4) - resolving polyunsaturated fatty acid

332 - eicosatetraenoic acid and lipoxins. 133. (2) (2013). LC-MS/MS method for - derived peptideAc(2 :

541 , et al. - Trends Pharmacol Sci 548.

(2012). Food

Sci Transl Med Trends sation. sation. - 26). Pediatr Gut - grade J

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 691 690 689 688 687 686 685 684 683 682 681 680 679 678 677 676 675 674 673 672 671 670 669 668 667 666 665 664 663 662 661 660 659 658 657 656 655 654 653 652 698 697 696 695 694 693 692

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, dietto the very Zadravec D, Brolinson A,Fisher RM, CarneheimCsikasz C, Bertrand-Michel RI, J Pharmacol Rev Basic and Clinical Ph RD,Ye Boulay F,Wang JM, Dahlgren C, Gerard C, Parmentier M reperfusion injury. brain barrier Wu Y, Wang YP, Guo YeP, XH, Wang J, Yuan SY Allergy Drug Targets WallaceJL (2006). Nitric oxide,aspirin-triggered lipoxins and NO-aspirin ingastric protection. 13- functional consequences. Vollmar B, Menger MD (2011). Intestinal ischemia/reperfusion: microcirculatory pathology and inflammatory receptors. (LXA4)A4 and LXA4 stable analoguesare potent inhibitors acuteof inflammation: evidencefor anti Takano Fiore T, S,Maddox JF, Brady HR, Petasis NA, Serhan CN(1997). Aspirin-triggered 15-epi-lipoxin 357 resolving properties of benzo Sun YP, Tjonahen E, KeledjianZhu R, M, Yang R, Recchiuti A Pathol Serhan CN,Yacoubian S,Yang(2008). R Anti-inflammatory and proresolving lipid mediators. 6 Serhan CN,Savill J (2005). Resolution inflammation:of the beginningthe programsend. and biological activities). Serhan CN,Samuelsson(1988). B Lipoxins: a new seriesof eicosanoids (biosynthesis,stereochemistry, formed from arachidonicacid inhuman leukocytes. Serhan CN,Hamberg M, Samuelsson B (1984). Lipoxins: novel series biologicallyof active compounds 158 neutrophils: evidencefor differe Serhan CN(1989). relationshipOn the between leukotrieneand lipoxin productionhuman by Opin Pharmacol This article isprotectedbycopyright. Allrights reserved. Schwab JM,Serhan CN(2006). Lipoxins and newlipid mediators inresolution the inflammation. of (12) 29. - - 366. 168. :

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal 1191 - 3: induced obesity. FASEB J -

long 279

Accepted Article- 1197. dysfunction and reduces MMP - - 312. chain fatty acid elongase

61(2)

6 (4)

J Mol Neurosci armacology. LXXIII. Nomenclaturefor formyl the peptide receptor (FPR)family.

: : 5

414 119 910-923. ,DOI :10.1111/bph.12957 (2) J Exp Med : Adv Exp BiolMed Comment citer cedocument: - -

Langenbeck'sarchives of surgery / Deutsche Gesellschaftfur Chirurgie 133 420. 161. - lipoxin A(4)analogs. -

137.

ntial metabolism of 15 24

46(3)

(11)

185

: (9) ELOVL3 in mice leadsto constrained lipid storage and resistance :

4366 483 :

229: 1693 - - 9 expression inarat model of focal cerebral ischemia 491. - 4377. , al.et

1 - 1704. -

14. ProstaglandinsLeukot EssentFatty Acids 29

Proc Natl Acad U Sci A S (2012). Alipoxin A4 analog amelioratesblood-

- HETE and 5 , et al. , et , etal. (2009). Anti-inflammatory and pro- - HETE. (2009). International Union of BiochimBiophys Acta

81 , etal. (17)

5335

(2010). Ablation of NatImmunol - 5339.

81(5 Annu Rev Inflamm

1004 396 - 6) - Curr - :

(1) (2)

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 703 702 701 700 699 705 704

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, (w/w) 5% fat of 2: polyunsaturatedTable Composition % the in acid3isocaloric of fatty diets containing Fish Oil Oil Colza Grape Oil Seed Vitamins Minerals Sucrose Methionin Starch Casein Cellulose This article isprotectedbycopyright. Allrights reserved. of 1: theTable Composition 3isocaloric % in diets

n - C22:6n C20:5n C18:3n C18:2n C18:1n 6/n C18:0 C16:0 C14:0 C12 N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal -

3 ratio

- - - - -

Accepted3 3 3 6 9 Article

Control 47.4 38.8 0.0 0.0 5.2 2.5 6.0 0.0 0.0 10 910-923. ,DOI :10.1111/bph.12957 Control

Comment citer cedocument: 21.8 43.6

2.5 2.5 1.0 4.5 0.2 22 0 2

70.0 18.4 n 0.0 0.0 1.0 4.2 6.5 0.0 0.0 70 - n 6

21.8 43.6

6 1.0 4.5 0.2

22 0 0 5 2

0.72 17.0 19.9 47.4 n 3.0 7.6 0.7 4.5 0.0 0.0 n - - 21.8 43.6 3 3 1.0 4.5 0.2 30

1 4 0 2

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 711 710 709 708 707 706

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, Total FA Total PUFA Total MUFA Total SAFA 20:4 n 22:6 n 22:5 n 22:4 n 22:2 n 22:5 n 20:5 n 20:4 n 20:3 n 20:3 n 20:2 n 18:3 n 18:3 n 18:2 n 24:1 n 22:1 n 20:1 n 18:1 n 18:1 n 17:1 n 16:1 n 16:1 n 15:1 n 14:1 n 22:0 21:0 20:0 18:0 16:0 15:0 14:0 12:0 10:0 acid. α fattyMUFA:acid;LA: linoleic acid; monounsaturated PUFA:acid; fatty ALA: polyunsaturated are n=10. diet express ±SEM, *p<0.05group. asmean vs.control satura SAFA: to9weekssubmitted ofcontrol, n This article isprotectedbycopyright. Allrights reserved. 3: acidConcentration (nmol/mgTable fatty of protein) inmouse intestine of - linolenic acid; AA: arachidoniclinolenic acid; AA:acid; EPA:

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal ------3 (DHA) 6 6 6 3 3 (EPA) 6 (AA) 6 3 6 3 (ALA) 6 6 (LA) 9 9 9 7 9 7 7 9 5 5 6 /22:6n

Accepted Article

- 3

910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument: 39.95 ±2.34 18.29 ±1.03 0.00 3.41 ±1.50 0.00 0.00 84.56 ±18.38 0.00 0.00 0.00 100.73 ±17.17 413.22 ±68.93 0.00 150.83 ±13.09 0.00 0.00 0.00 0.00 0.00 0.00 185.97 ±23.68 286.85 ±43.95 0.00 0.00 0.00 0.00 Control 1284 ±151 148.08 ±24.46 664.78 ±99.19 472.82 ±67.64 9.51 ±0.75 1.88 ±1.21 0.00 0.00 0.00 0.00 0.00

- 3 orn

- 6 diet before fatty acid (FA) metabolite extraction.fatty(FA) Data metabolite 6 dietbeforeacid 0.00 0.00 0.00 0.00 147.24 ±21.21 349.08 ±75.12 0.00 5.39 ±2.57 0.00 0.00 n 1345 ±162 144.50 ±19.93 700.33 ±99.38 501.71 ±98.91 0.73 ±0.07 19.87 ±3.14 0.00 0.00 0.00 5.69 ±0.78 10.78 ±1.86 13.81 ±1.19 8.56 ±0.86 0.00 0.00 15.65 ±1.65 0.00 69.76 ±10.07 0.00 0.00 0.00 93.00 ±11.58 450.11 ±49.71 0.00 157.22 ±38.09 0.00 0.00 - 3

eicosapentaenoicacid; docosahexaenoic DHA:

* 31

Diet

* * * * * * *

0.00 0.00 0.00 0.00 127.53 ±42.45 329 ±79.41 0.00 1.02 ± 0.00 0.00 n 1432 ±114 290.78 ±32.30 685.58 ±99.80 457.98 ±122.88 44.09 ±11.34 0.93 ±0.52 0.00 0.00 0.00 0.00 0.00 60.37 ±5.70 13.47 ±1.34 0.00 6.97 ±0.92* 0.00 0.00 209.05 ±23.64 0.00 0.00 0.00 94.35 ±19.73 445.18 ±65.70 0.00 146.05 ±14.37 0.00 0.00 - 6

* *

ted fatty acid;ted fatty . Mice were

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 730 729 728 727 726 725 724 723 722 721 720 719 718 717 716 715 714 713 712 734 733 732 731

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, spectrometry in sham ischemia (white bar), sham I/R animals (stripped bar) and following 50 50 following and bar) (stripped animals I/R sham bar), (white ischemia sham in spectrometry A lipoxin of Synthesis ( clusters. the represents line red dotted The metabolites. PUFA the of correlation the reflects a indicates color green quantity a row, the of mean the than greater quantity a indicates color red A column). (in mice of groups the and row) (in metabolites PUFA different the for obtained scores chromatography (A) diets. different three the to and I/R to miceexposed of intestine in metabolites PUFA of Quantification 3: Figure sham; + significan p<0.05, p<0.01, ++ 6 n= SEM, ± mean n bar), (white control to exposed sham, ( weeks. n control, to exposed mice bar) (black I/R or bar) (white sham in ( scores damage microscopic followed: were parameters n control, receiving and I/R to submitted eosin I/R. by induced intestine the of Inflammation 2: Figure group. significantly ***p<0.001, grou from **p<0.01, different control diet n control, to submitted composition. n acids (C20:4 fatty acid hepatic arachidonic the on intervention dietary of Effect 1: Figure Figure legends This article isprotectedbycopyright. Allrights reserved. - N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal tie hsooia scin o sal netn fo sham from intestine small of sections histological stained

D lower than the mean and black color a quantity close to the mean. The dendrogram dendrogram The mean. the to close quantity a color black and mean the than lower

Accepted- Article E ) CXCL1 and CCL2 expressions were quantified in quantified were expressions CCL2 and CXCL1 ) - tandem mass spectr tandem - 8 per group. * p <0.05, ** p<0.01, *** p<0.001, significantly different from from different significantly p<0.001, *** p<0.01, ** <0.05, p * group. per 8 910-923. ,DOI :10.1111/bph.12957 - Comment citer cedocument: 3 or n or 3

- Heat (LxA ) n dcshxeoc cd C26 n (C22:6 acid docosahexaenoic and 6) - 6 PUFA diet for 9 weeks. Values are mean ± SEM, n= 6 n= SEM, ± mean are Values weeks. 9 for diet PUFA 6 - map and clustering of quantified PUFA metabolites by liquid liquid by metabolites PUFA quantified of clustering and map 4 ) was measured by liquid liquid by measured was )

ometry. The heatmapaverage coding acolor of ometry. represents The z tly different from control diet group. from control different tly diet - 3 (black bar) or n or bar) (black 3

- o n or 3 32

- PF de. Different diet. PUFA 6 B ), myeloperoxidase activity (MPO) ( (MPO) activity myeloperoxidase ), -

( 6 (gray bar) PUFA diet. Values are are Values diet. PUFA bar) (gray 6 A ) representative hematoxylin and and hematoxylin representative ) chromatography I/R mice mice I/R - - prtd os o mouse or mouse operated - o n or 3 ) ai i lvr f mice of liver in ratio 3)

versus p. - 6 PUFA diet for 9 9 for diet PUFA 6

- corresponding adm mass tandem inflammatory inflammatory - 8 per per 8 Total C B - ) )

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 754 753 752 751 750 749 748 747 746 745 744 743 742 741 740 739 738 737 736 735 757 756 755

J., Dubourdeau, M.,Guillou, H.,Perretti, M.,Vergnolle, N.(Auteur decorrespondance), Cenac, Gobbetti, T.,Ducheix, S.,leFaouder, P.,Perez,T., Riols,F.,Boueé, J.,Bertrand-Michel, relevant to the manuscript. relevant themanuscript. to Disclosures: ++I/Rdifferent p<0.0from1, significantly group. vehicle 6 n= SEM, ± mean are Values group. diet control in effect induced I/R of inhibition of percentage expo and i.v.) ischemia before µg/kg I/Rmetabolites insham(grey (blackmicebar) treated orantagonist withFPR2 bar) (BOC n to exposed and i.v.) ischemia before µg/kg treatedmice I/Rsham(blackbar) (greyor in bar) expression, (C chemokine scores, damage microscopic activity, myeloperoxidase followed: were parameters inflammatory (A) PUFA dieteffects. anti-inflammatory n-6 FPR2 in Figureantagonist Role of 5: differentfromgroup. corresponding ischemia 6 n= SEM, ± mean are Values µm). 50 (scale: intestine small the in detected immunoreactivity n or I/Rquantified mice in intestine small mouse in FPR2 of Expression 4: Figure significantly ischemiafrom control different diet group per group. animals 8 to 6 of SEM ± means represent and protein pg/mg in expressed are Data bar). black (I/R, reperfusion of hours 5 and ischemia of minutes 50 or bar) (gray ischemia of minutes This article isprotectedbycopyright. Allrights reserved. - 8 per group. * p <0.05, ** p<0.01, *** p<0.001, significantly different from sham; + p <0.05, p + sham; from different significantly p<0.001, *** p<0.01, ** <0.05, p * group. per 8 - (ry a) UA it ( diet. PUFA bar) (gray 6 N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal

Accepted Article p<0.01, ++ sham; corresponding from different significantly p<0.001, *** p<0.01, **

The authors have no potential conflicts (financial,orpersonal)conflicts have that are authors professional, nopotential The - X - mtf lgn 1 CC1 epeso ad hmkn lgn 2 (CCL2) 2 ligand chemokine and expression (CXCL1) 1 ligand motif) C 910-923. ,DOI :10.1111/bph.12957 versus Comment citer cedocument:

corresponding sham, exposed to control (white bar),n (white tocontrol exposed corresponding sham, B

Pcue o FR, yoeai 1, D5 n Ly and CD45 18, cytokeratin FPR2, of Pictures ) sed to n to sed

- - 6 PUFA diet for 9 weeks. ( weeks. 9 for diet PUFA 6 6 PUFA diet for 9 weeks. Data are expressed as expressed are Data weeks. 9 for diet PUFA 6 33 - 8 per group. * p <0.05, ** p<0.01, significantly p<0.01, ** <0.05, p * group. per 8 of mice exposed to sham and to n to and sham to exposed mice of

( . with FPR2 antagonist(BOC FPR2 with A ) P2 RA xrsin was expression mRNA FPR2 B

) N ) - 6 PUFA derived derived PUFA 6 - - 6 PUFA diet PUFA 6 3 (black bar) (blackbar) 3 Different Different - - 2, 500 2, 500 500 2, - 6B.2 6B.2 Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 759 758

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument:

bph_12957_f1 34

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 761 760

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument:

bph_12957_f2 35

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 763 762

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument:

bph_12957_f3 36

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 765 764

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument:

bph_12957_f4 37

Version postprint ischaemia/reperfusion injury involve lipoxin A4 and its receptor. British Journal of Pharmacology, 172 (3), 767 766 View publication stats

N. (2015). Protective effects ofn-6 fattyacids-enriched dieton intestinal Accepted Article 910-923. ,DOI :10.1111/bph.12957 Comment citer cedocument:

bph_12957_f5 38