Journal of Functional Foods 17 (2015) 575–586 Available online at www.sciencedirect.com ScienceDirect journal homepage: www.elsevier.com/locate/jff Intake of Passiflora edulis leaf extract improves antioxidant and anti-inflammatory status in rats with 2,4,6-trinitrobenzenesulphonic acid induced colitis Cinthia Baú Betim Cazarin a, Juliana Kelly da Silva a, Talita Cristina Colomeu b, Ângela Giovana Batista a, Laura Maria Molina Meletti c, Jonas Augusto Rizzato Paschoal a, Stanislau Bogusz Junior d, Patrícia Aparecida de Campos Braga a, Felix Guillermo Reyes Reyes a, Fábio Augusto e, Luciana Rodrigues de Meirelles b, Ricardo de Lima Zollner b,*, Mário Roberto Maróstica Júnior a,** a School of Food Engineering, University of Campinas, Rua Monteiro Lobato, 80, Cidade Universitária Zeferino Vaz, Campinas, São Paulo, Brazil b Faculty of Medical Science, University of Campinas, Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, Campinas, São Paulo, Brazil c Center of Research and Development on Plant Genetic Resources, Agronomic Institute of Campinas, Campinas, SP, Brazil d Institute of Chemistry of São Carlos (IQSC), University of São Paulo (USP), São Carlos, São Paulo, Brazil e Institute of Chemistry, Instituto Nacional de Ciência e Tecnologia em Bioanalítica (INCTBio), University of Campinas, Campinas, São Paulo, Brazil ARTICLE INFO ABSTRACT Article history: The antioxidant and anti-inflammatory effects of the aqueous extract of Passiflora edulis leaves, Received 18 December 2014 which is a source of vitexin, isovitexin and isoorientin, was investigated in a 2,4,6- Received in revised form 27 May trinitrobenzenesulphonic acid induced colitis model. The oral intake of P. edulis extract 2015 (1100 µgmL−1) improved significantly (P < 0.05) the endogenous antioxidant status and de- Accepted 28 May 2015 creased lipid peroxidation in serum, liver, and colon. The consumption of P. edulis extract Available online 24 June 2015 decreased the level of pro-inflammatory in colon tissue, especially by reducing IL-1β five- fold and TNF-α two-fold compared to the control colitis group. The extract of P. edulis leaves Keywords: possessed therapeutic activity in healthy rats and in TNBS-induced colitis, especially by con- Passiflora edulis leaves trolling the oxidative stress and inflammation observed in this disease. Colitis © 2015 Elsevier Ltd. All rights reserved. Oxidative stress Antioxidant potential Anti-inflammatory activity * Corresponding author. Faculty of Medical Science, University of Campinas, Rua Tessália Vieira de Camargo, 126, Cidade Universitária Zeferino Vaz, Campinas, São Paulo, Brazil. Tel.: +55 19 32893709; fax: +55 19 32893709. E-mail address: [email protected] (R.L. Zollner). ** Corresponding author. School of Food Engineering, University of Campinas, Rua Monteiro Lobato, 80, Cidade Universitária Zeferino Vaz, Campinas, São Paulo, Brazil. Tel.: +55 19 3521 4059; fax: +55 19 3521 4059. E-mail address: [email protected] (M.R. Maróstica Júnior). http://dx.doi.org/10.1016/j.jff.2015.05.034 1756-4646/© 2015 Elsevier Ltd. All rights reserved. 576 Journal of Functional Foods 17 (2015) 575–586 thus modulating the production of short-chain fatty acids 1. Introduction (SCFA). The improvement in the mucosal protection against in- flammatory damage has been observed in many studies in Passiflora is a native tropical and subtropical species culti- which the formation of SCFA increased after the consump- vated in several parts of the world, especially in the American tion of microbiota substrates (Duda-Chodak, 2012; Parkar et al., continent, and encompasses approximately 450 species, usually 2013). Therefore, phenolic compounds may play a role in im- called passion fruit (Dhawan, Dhawan, & Sharma, 2004). Their proving intestinal health (Duda-Chodak, 2012; Parkar et al., cultivation is primarily focused on the production of pulp juice 2013). and frozen pulp from the fruit. However, in folk medicine, leaves Inflammatory bowel disease (IBD) is a chronic immune- of Passiflora are used due to their anxiolytic (Li et al., 2011) and mediated intestinal disorder that encompasses Crohn’s disease anti-inflammatory activities (Montanher, Zucolotto, Schenkel, (CD) and ulcerative colitis (UC). CD is a transmural inflamma- & Frode, 2007), which may be mainly associated with the pres- tion that can affect the entire gastrointestinal tract, while UC ence of phenolic compounds (lucenin-2, vicenin-2, isoorientin, is a non-transmural inflammation restricted to the colon, and isovitexin, luteolin). In a previous work, we showed that Passiflora there are many factors related to colitis pathogenesis, includ- alata extract contained seven-fold more of the total amounts ing genetic, environmental, microbiological and immunological of flavonoids vitexin, isovitexin, and isoorientin than Passiflora factors (Podolsky, 2002). edulis extract (Colomeu et al., 2014; da Silva et al., 2013). Some Increases in reactive oxygen species (ROS) in colorectal evidence about the anti-inflammatory activity of P. alata was biopsy specimens of subjects and animal models of IBD are observed by Colomeu et al. (2014) in a type 1 diabetes experi- also observed (Keshavarzian et al., 1992; Nieto et al., 2000). The mental model. Intake of aqueous extract of P. alata promoted cause of the oxidative stress is an imbalance between protec- a decrease in cell infiltration and insulitis in NOD mice com- tor systems and production of free radicals. Our previous data pared to the control group. However, the amounts of isovitexin indicate that the extract of P. edulis leaves possessed antioxi- in P. edulis extract (0.50 ± 0.04 mg g−1) were almost three-fold dant activity in vitro and in vitro against oxidative damage, and more than that in P. alata extract (0.19 ± 0.01 mg g−1)(Colomeu is considered as an important source of natural antioxidants et al., 2014; da Silva et al., 2013). Protection against oxidative (da Silva et al., 2013). In addition, we demonstrated that it could stress was observed in cells treated with isovitexin extract from reduce oxidative stress in vivo by improving the antioxidant rice Oryza sativa, mainly through decreased lipid peroxidation power, reducing TBARS (thiobarbituric acid reactive sub- and inhibition of xanthine oxidation activity (Lin, Chen, Lee, stances) level, and by increasing the number of useful colonic & Lin, 2002). Amongst the flavonoids identified in the aqueous microbiota in healthy rats (da Silva et al., 2013). In the face of extract of P. edulis, isoorientin showed a higher amount this, the aim of the present study was to evaluate the effect (1.05 ± 0.03 mg g−1) compared with others flavonoids (da Silva of intake of aqueous P. edulis extract on inflammation and oxi- et al., 2013). dative stress using an experimental model of rat colitis induced The administration of hydroalcoholic extract of P. edulis in by 2,4,6-trinitrobenzenesulphonic acid (TNBS). rat colon anastomosis showed an anti-inflammatory effect through increased infiltration of mononuclear cells, fibro- blast proliferation, and granulation tissue, promoting the colon 2. Materials and methods healing (Bezerra et al., 2006). In another study, aqueous ex- tracts of leaves of P. alata (100 mg kg−1, i.p.) and P. edulis 2.1. P. edulis leaves (250 mg kg−1, i.p.) showed a significant anti-inflammatory ac- Leaves of P. edulis were collected in June 2011 from the Agro- tivity on carrageenan-induced pleurisy in mice, especially by nomic Institute of Campinas (Campinas, SP, Brazil). A voucher inhibition of influx of leukocytes and a reduction in the serum specimen was deposited in the Herbarium at the same insti- C-reactive protein levels (Vargas et al., 2007). tute. The leaves were dried in an oven with circulating air In addition, different aqueous extract sources of polyphe- (Marconi, Piracicaba, SP, Brazil) at 50 °C for 48 h, ground to a nols have shown anti-carcinogenic activity (de Mejía, Song, fine homogeneous powder, and stored in amber glass bottles Heck, & Ramírez-Mares, 2010; Li et al., 2013; Zhao et al., 2014). at 8 °C. The anti-carcinogenic activity from P. edulis was evaluated in the fruit and the peel showed 89% inhibition of HT-29 (human 2.2. Aqueous P. edulis leaf extract – Tea colon adenocarcinoma cells) cancer cell line activity, and the pulp and seed showed the highest inhibitory activity (97%) (Li The extract was prepared by infusing powdered leaves in boiling et al., 2013). water for 25 min (leaves/water, 1:25, w/v). The extract was fil- These aforementioned effects may be correlated with the tered, and the volume adjusted to 25 mL. The antioxidant polyphenols found in the extracts, due to their antioxidant prop- potential and the composition of phenolic compounds were erties and ability to neutralize or quench oxidants (Bezerra et al., analysed in the extract. The concentration (1100 µgmL−1)ofthe 2006; Vargas et al., 2007). Thus, some hypotheses about the poly- aqueous extract (named Tea) given to the animals was deter- phenol metabolism have been studied, especially because, mined in accordance to a previous study (da Silva et al., 2013). before their absorption, most dietary polyphenols are trans- formed in the colon by microbiota, which may influence their 2.3. LC–MS/MS analysis absorption and biological activities (Parkar, Trower, & Stevenson, 2013). Unabsorbed phenolic compounds that could remain in The LC–MS/MS equipment used in this study for the identifi- the intestinal lumen could be metabolized by the microbiota, cation of the flavonoids in P. edulis aqueous extract consists