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Research Article Involvement of the PI3K/Akt/NF- B Hindawi BioMed Research International Volume 2017, Article ID 9698410, 13 pages https://doi.org/10.1155/2017/9698410 Research Article Involvement of the PI3K/Akt/NF-휅B Signaling Pathway in the Attenuation of Severe Acute Pancreatitis-Associated Acute Lung Injury by Sedum sarmentosum Bunge Extract Yuepeng Jin,1 Lewei Liu,2 Bicheng Chen,3 Yongyu Bai,4 Fan Zhang,4 Qiang Li,4 Chongqing Lv,5 Hongwei Sun,1 Junjian Li,1 Sadman Rubby,4 Lihong Yang,1 Roland Andersson,6 and Mengtao Zhou1 1 Department of Surgery, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang Province, China 2YueQing Affiliated Hospital of Wenzhou Medical University, YueQing People’s Hospital, Yueqing, Zhejiang Province, China 3Zhejiang Provincial Top Key Discipline in Surgery, Wenzhou Key Laboratory of Surgery, Department of Surgery, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang Province, China 4Wenzhou Medical University, Wenzhou, Zhejiang Province, China 5Shengli Oilfield Central Hospital, Dongying, Shandong Province, China 6Department of Surgery, Clinical Sciences Lund, Lund University and Lund University Hospital, Sweden Correspondence should be addressed to Mengtao Zhou; [email protected] Received 21 July 2017; Revised 19 September 2017; Accepted 22 October 2017; Published 5 December 2017 Academic Editor: Fabrizio Montecucco Copyright © 2017 Yuepeng Jin et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Sedum sarmentosum Bunge possesses excellent anti-inflammatory properties and was used in the treatment of inflammatory diseases. The aim of the present study was to investigate the efficiency of Sedum sarmentosum Bunge extract (SSBE) on severe acute pancreatitis-associated (SAP-associated) acute lung injury (ALI) in rats and to explore the underlying mechanisms. Here, we used a sodium taurocholate-induced SAP rat model to determine the role of SSBE in ALI. During the course of pancreatitis, the expressions of phosphorylated phosphoinositide 3-kinases (PI3K)/protein kinase B (Akt) and nuclear factor-kappa B (NF-B) p65 in the lungs were upregulated. Meanwhile, a parallel increase in the levels of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor- (TNF-) in the lungs was observed after the induction of SAP. Treatment with SSBE significantly reduced the expression of p-Akt and p-p65 in the lungs and attenuated the severity of SAP-associated ALI compared to the SAP group at 12 h and 24 h. In summary, this study showed that SSBE has beneficial effects on SAP-associated ALI, probably through the PI3-K/Akt signalingpathwaysbysuppressingtheNF-Bactivities. 1. Introduction contributes to early mortality (i.e., during the first week) [5, 8, 11–14]. Severe acute pancreatitis is a potentially lethal inflamma- Although there are numerous factors involved in the torydiseasewithvariableinvolvementoftheperipancreatic development of SAP-associated ALI, inflammatory medi- organs, and/or remote organ systems, by developing a sys- ators have been reported to play a dominant role in the temic inflammatory response syndrome (SIRS) [1–7]. Among pathogenesis of SAP-associated ALI [7, 15]. Activation of the systemic complications, ALI is the most frequent and inflammatory signaling pathways, such as PI3-K/Akt and potentially the most serious and may occur in 15% to 55% NF-B, is involved as underlying molecular mechanisms in of SAP cases, including both ALI and the acute respiratory thedevelopmentofALI[7,16–19].Theupstreamregulator distress syndrome (ARDS) [8–10]. The associated mortality of NF-B involves PI3-K/Akt [16] signaling pathways. The rate in SAP has been reported to be from 10% and up, where underlyingmechanismsofSAP-associatedALIarethuscom- respiratory dysfunction (ALI and ARDS) is what mostly plex, which may explain the lack of specific pharmacological 2 BioMed Research International interventions, despite innumerous attempts over the years 2.5. The Animal Model for Induction of SAP. All procedures [14, 20–22]. wereperformedinsterileconditions.Theratswereanes- Sedum sarmentosum Bunge is a perennial succulent herb thetized by subcutaneous injection of 10% chloral hydrate frequently used in the treatment of inflammatory diseases solution (0.3 mL/100 g). According to Aho et al. [32], SAP in Oriental countries [23–25]. The complete genome of was induced by retrograde injection of 5% sodium tauro- Sedum sarmentosum has been sequenced [26] and previ- cholate (1 ml/kg) into the main pancreatic duct through a ous studies have revealed that Sedum sarmentosum Bunge microinjection pump at a speed of 0.1 ml/min. The control possess anti-inflammatory, antiangiogenic, antinociceptive, group had the same surgical procedure, but without the and antifibrogenic properties [27–29]. Synthesis of mono- cannulation procedure and the retrograde injection of 5% cyte and macrophage-mediated inflammatory factors can be sodium taurocholate. decreased by Sedum sarmentosum Bunge [29]. Active chem- ical components in Sedum sarmentosum Bunge identified 2.6. Preparation and Administration of Sedum sarmentosum include megastigmane glycoside, quercetin, isorhamnetin, Bunge Extract. The extract protocol of Sedum sarmentosum and kaempferide [27, 30]. Sedum sarmentosum possess anti- Bunge has been shown by Bai et al. [29]. The whole plants inflammatory properties, and side effects seem to be lim- were soaked in the tenfold distilled water for one hour and ited [28]. In a previous preliminary study, we found anti- were used in the refluxing process of two hours. The extract inflammatory effects and an attenuation of the morphological was then filtrated and concentrated. Moreover, the drugs of injury in the lungs in a rat model of SAP [31]. the refluxing extract need further refluxing, filtration, and The purpose of the present study was to verify whether concentration. For animal experiments, 1 g of SSBE was dis- SSBE attenuated the severity of SAP-associated ALI. If this solved in 10 ml of normal saline, resulting in a concentration was the case, we aimed at elucidating the underlying mecha- of 100 mg/ml. nisms explaining the effect of SSBE in ALI. SSBE at a dose of 100 mg/kg was immediately adminis- tered subcutaneously and every 12 h following the establish- 2. Materials and Methods ment of the SAP model in the SSBE group. The 3, 6, and 12 h 2.1. Animals. The Laboratory Animal Center of Wen- subgroups in the SSBE group were all administered with SSBE zhou Medical University (Wenzhou, China) supplied 84 once. The 24 h subgroup in the SSBE group was administered healthy male Sprague-Dawley (SD) rats (weighing 250–300 g, with SSBE two times. Normal saline was administered in the 10–12 weeks old). The rats were maintained under specific same way as the other two groups. pathogen-free(SPF)conditionsandthetemperaturewaskept ∘ 2.7. Collection of Specimens. 3, 6, 12, and 24 h after the at 20–22 C. A 12 h light-dark cycle was maintained, and all induction of SAP, seven rats from each group were randomly the rats were fed with a standard rat diet and water. All selected to undergo abdominal incision under anesthesia. experiments were performed on animals after 12 h of fasting, Blood and pancreatic and lung tissues were collected for but with free access to water. further analysis. 2.2. Ethics Statement. The Institutional Animal Committee of Wenzhou Medical University, Wenzhou, China, approved 2.8. Histopathological Analysis. Tissues from the pancreas the protocol for the animal experiment (Permit Number: and lungs were harvested and fixed in 10% formaldehyde wydw2013-0002). All animals received care in accordance solution, embedded in paraffin, sectioned, and stained with with “Guide for the Care and Use of Laboratory Animals.” All hematoxylin-eosin (H&E) for light microscopy. The pancreas surgical interventions were performed under chloral hydrate pathological grading described by Schmidt et al. [33] was anesthesia. used, considering all factors, including edema, acinar necro- sis, hemorrhage, fat necrosis, inflammation, and perivascular 2.3. Animals and Groups. Eighty-four male SD rats were infiltration, when scoring the pancreatic injury in experimen- randomly divided into 3 groups: control group (control, = tal SAP.Lung pathological scores were evaluated according to 28), SAP group (SAP, =28), and SSBE treatment group the method reported by Osman et al. [34]. An experienced (SSBE, =28). Every group was divided into 4 subgroups, pathologist examined the pathological sections unaware of namely,3,6,12,and24hsubgroups(=7animals/group). the treatment groups, that is, in a blinded fashion. 2.4. Materials. The following materials had been purchased 2.9. Serum Amylase Activity and Levels of IL-1,IL-6, from the sources as mentioned below: Sedum sarmentosum and TNF- in Lung Tissue. Serum amylase activity was Bunge extract (Xuancheng Baicao Plant Industry and Trade determined by means of iodine-amylum colorimetry and Co., Ltd., Anhui, China); sodium taurocholate (Sigma Chem- expressed in U/L. Lung tissue was homogenized and cen- ical Company, St Louis, MO, USA); Rat ELISA Kit for detect- trifuged, and the supernatants expressed in pg/ml were col- ing IL-1,IL-6,andTNF- (RapidBio Company, CA, USA); lected to detect IL-1,IL-6,andTNF- by means of enzyme- MPO Kit (JianCheng Biotechnology Research Institute, Nan- linked immunosorbent assay (ELISA) using kits provided by jing, China);
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