Protective effects of Semen Crotonis Pulveratum on trinitrobenzene sulphonic acid-induced colitis in rats and H2O2-induced intestinal cell apoptosis in vitro

Wang,Xiaohong; Zhao,Jie; Han,Zhe; Tang,Fang

doi:10.3892/ijmm.2015.2175

Protective effects of Semen Crotonis Pulveratum on trinitrobenzene sulphonic acid-induced colitis in rats and H2O2-induced intestinal cell apoptosis in vitro

Authors:
- Xiaohong Wang
- Jie Zhao
- Zhe Han
- Fang Tang
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Affiliations: Tianjin Medical University General Hospital, Tianjin 300052, P.R. China, Basic Medical College, Tianjin Medical University, Tianjin 300070, P.R. China
Published online on: April 7, 2015 https://doi.org/10.3892/ijmm.2015.2175
Pages: 1699-1707

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Abstract

Ulcerative colitis (UC) is a chronic inflammatory bowel disease. Semen Crotonis Pulveratum (SCP) has been used as a traditional medicine for the treatment of UC. However, its molecular mechanisms of action have not yet been elucidated. In the present study, we aimed to investigate the preliminary mechanisms of the role of SCP on trinitrobenzene sulphonic acid (TNBS)-induced UC in rats and hydrogen peroxide (H2O2)-induced intestinal cell apoptosis in vitro. Wistar rats (n=9 per group) were randomly divided into 4 groups: the normal control group, the UC group, the UC + SCP group and the UC + sulfasalazine group as a positive control. The proportion of CD4+CD25+ T cells and CD4+CD25+Foxp3+ Tregs, and the expression levels of interleukin (IL)-6 and IL-10 in the peripheral blood, as well as the expression levels of cyclooxygenase-2 (COX-2) and intercellular adhesion molecule-1 (ICAM-1) in the colon tissues were determined by flow cytometry, ELISA and immunohistochemical staining, respectively. Rat intestinal epithelial (IEC-6) cell apoptosis induced by H2O2 was determined by TUNEL assay, flow cytometry using Annexin V/propidium iodide (PI) staining and western blot analysis of caspase-3 activation, respectively. Significantly higher proportions of circulating CD4+CD25+ T cells and CD4+CD25+Foxp3+ Tregs were present in the UC + SCP group compared with the UC group. A significantly decreased expression of IL-6 and an increased expression of IL-10 were also observed in the UC + SCP group compared with UC group. SCP significantly reduced the UC-induced increase in the expression of COX-2 and ICAM-1 in the colon tissues. SCP inhibited cell apoptosis and caspase-3 activation induced by H2O2 in the ICE-6 cells. Our data thus indicate that SCP inhibits inflammation in UC by increasing the proportion of circulating Tregs, altering cytokine production and decreasing COX-2 and ICAM-1 expression. In addition it protects against H2O2-induced intestinal cell apoptosis in vitro.

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1	Burisch J, Pedersen N, Čuković-Čavka S, et al: EpiCom-group: East-West gradient in the incidence of inflammatory bowel disease in Europe: The ECCO-EpiCom inception cohort. Gut. 63:588–597. 2014. View Article : Google Scholar
2	Radwan P, Radwan-Kwiatek K, Tabarkiewicz J, Radej S and Rolinski J: Enhanced phenotypic and functional maturation of monocyte-derived dendritic cells from patients with active Crohn's disease and ulcerative colitis. J Physiol Pharmacol. 61:695–703. 2010.
3	Zhao QJ, Yu YB, Zuo XL, Dong YY and Li YQ: Milk fat globule-epidermal growth factor 8 is decreased in intestinal epithelium of ulcerative colitis patients and thereby causes increased apoptosis and impaired wound healing. Mol Med. 18:497–506. 2012.
4	Serrano D, Bhowmick T, Chadha R, Garnacho C and Muro S: Intercellular adhesion molecule 1 engagement modulates sphingomyelinase and ceramide, supporting uptake of drug carriers by the vascular endothelium. Arterioscler Thromb Vasc Biol. 32:1178–1185. 2012. View Article : Google Scholar : PubMed/NCBI
5	Kim YJ, Lee JS, Hong KS, Chung JW, Kim JH and Hahm KB: Novel application of proton pump inhibitor for the prevention of colitis-induced colorectal carcinogenesis beyond acid suppression. Cancer Prev Res (Phila). 3:963–974. 2010. View Article : Google Scholar
6	Flannigan KL, Agbor TA, Blackler RW, et al: Impaired hydrogen sulfide synthesis and IL-10 signaling underlie hyperhomocysteinemia-associated exacerbation of colitis. Proc Natl Acad Sci USA. 111:13559–13564. 2014. View Article : Google Scholar : PubMed/NCBI
7	Yu QT, Saruta M, Avanesyan A, Fleshner PR, Banham AH and Papadakis KA: Expression and functional characterization of FOXP3⁺ CD4⁺ regulatory T cells in ulcerative colitis. Inflamm Bowel Dis. 13:191–199. 2007. View Article : Google Scholar : PubMed/NCBI
8	Imaoka A, Shima T, Kato K, Mizuno S, Uehara T, Matsumoto S, Setoyama H, Hara T and Umesaki Y: Anti-inflammatory activity of probiotic Bifidobacterium: Enhancement of IL-10 production in peripheral blood mononuclear cells from ulcerative colitis patients and inhibition of IL-8 secretion in HT-29 cells. World J Gastroenterol. 14:2511–2516. 2008. View Article : Google Scholar : PubMed/NCBI
9	Chen X: Protective effects of quercetin on liver injury induced by ethanol. Pharmacogn Mag. 6:135–141. 2010. View Article : Google Scholar : PubMed/NCBI
10	Liu DY, Guan YM, Zhao HM, Yan DM, Tong WT, Wan PT, Zhu WF, Liu HN and Liang XL: The protective and healing effects of Si Shen Wan in trinitrobenzene sulphonic acid-induced colitis. J Ethnopharmacol. 143:435–440. 2012. View Article : Google Scholar : PubMed/NCBI
11	Lindsay JO and Hodgson HJ: Review article: The immunoregulatory cytokine interleukin-10 - a therapy for Crohn’s disease. Aliment Pharmacol Ther. 15:1709–1716. 2001. View Article : Google Scholar : PubMed/NCBI
12	Kim SJ, Kim YG, Kim DS, et al: Oldenlandia diffusa ameliorates dextran sulphate sodium-induced colitis through inhibition of NF-κB activation. Am J Chin Med. 39:957–969. 2011. View Article : Google Scholar
13	Heinrich PC, Behrmann I, Müller-Newen G, Schaper F and Graeve L: Interleukin-6-type cytokine signalling through the gp130/Jak/STAT pathway. Biochem J. 334:297–314. 1998.PubMed/NCBI
14	Gracie DJ and Ford AC: Evidence-based management of ulcerative colitis. Minerva Gastroenterol Dietol. 58:87–99. 2012.PubMed/NCBI
15	Ford AC, Bernstein CN, Khan KJ, Abreu MT, Marshall JK, Talley NJ and Moayyedi P: Glucocorticosteroid therapy in inflammatory bowel disease: Systematic review and meta-analysis. Am J Gastroenterol. 106:590–599. 2011. View Article : Google Scholar : PubMed/NCBI
16	Prantera C and Marconi S: Glucocorticosteroids in the treatment of inflammatory bowel disease and approaches to minimizing systemic activity. Therap Adv Gastroenterol. 6:137–156. 2013. View Article : Google Scholar : PubMed/NCBI
17	Oz HS, Chen T and de Villiers WJ: Green tea polyphenols and sulfasalazine have parallel anti-inflammatory properties in colitis models. Front Immunol. 4:1322013. View Article : Google Scholar : PubMed/NCBI
18	Margagnoni G, Pagnini C, Menasci F, Festa S and Delle Fave G: Critical review of the evidence on 5-aminosalicilate for chemoprevention of colorectal cancer in ulcerative colitis: A methodological question. Curr Clin Pharmacol. 9:84–90. 2014. View Article : Google Scholar
19	Watson S, Pensabene L, Mitchell P and Bousvaros A: Outcomes and adverse events in children and young adults undergoing tacrolimus therapy for steroid-refractory colitis. Inflamm Bowel Dis. 17:22–29. 2011. View Article : Google Scholar
20	Kruis W, Schreiber S, Theuer D, et al: Low dose balsalazide (1.5 g twice daily) and mesalazine (0.5 g three times daily) maintained remission of ulcerative colitis but high dose balsalazide (3.0 g twice daily) was superior in preventing relapses. Gut. 49:783–789. 2001. View Article : Google Scholar : PubMed/NCBI
21	Minaiyana M, Ghannadib A, Mahzounic P and Jaffari-Shirazid E: Comparative study of Berberis vulgaris fruit extract and berberine chloride effects on acetic acid-induced colitis in rats. Iran J Pharm Res. 10:97–104. 2010.
22	Atyabi F, Vahabzadeh R and Dinarvand R: Preparation of ethylcellulose coated gelatin microspheres as a multiparticulate colonic delivery system for 5-aminosalicilic acid. Iran J Pharm Res. 2:81–86. 2004.
23	Zhang J, Gao W, Hu X, Liu Z and Liu C: The influence of compatibility of traditional Chinese medicine on the pharmacokinetic of main components in Fructus aurantii. J Ethnopharmacol. 144:277–283. 2012. View Article : Google Scholar : PubMed/NCBI
24	Chinese Pharmacopoeia Commission: Chinese Pharmacopoeia Commission: Pharmacopoeia of the People’s Republic of China. Chemical Industry Press; Beijing: 2005
25	Liu DY, Zhao HM, Zhao N, Xin ZP and Lu AP: Pharmacological effects of ba-wei-xi-lei powder on ulcerative colitis in rats with enema application. Am J Chin Med. 34:461–469. 2006. View Article : Google Scholar : PubMed/NCBI
26	Vilela EG, Torres HO, Martins FP, Ferrari ML, Andrade MM and Cunha AS: Evaluation of inflammatory activity in Crohn's disease and ulcerative colitis. World J Gastroenterol. 18:872–881. 2012. View Article : Google Scholar : PubMed/NCBI
27	Ślebioda TJ and Kmieć Z: Tumour necrosis factor superfamily members in the pathogenesis of inflammatory bowel disease. Mediators Inflamm. 2014:3251292014. View Article : Google Scholar : PubMed/NCBI
28	Boschetti G, Nancey S, Sardi F, Roblin X, Flourié B and Kaiserlian D: Therapy with anti-TNFα antibody enhances number and function of Foxp3(+) regulatory T cells in inflammatory bowel diseases. Inflamm Bowel Dis. 17:160–170. 2011. View Article : Google Scholar
29	Heller F, Florian P, Bojarski C, Richter J, Christ M, Hillenbrand B, Mankertz J, Gitter A, Burgel N and Fromm M: Interleukin-13 is the key effector Th2 cytokine in ulcerative colitis that affects epithelial tight junctions, apoptosis, and cell restitution. Gastroenterology. 129:550–564. 2005. View Article : Google Scholar : PubMed/NCBI
30	Kamikozuru K, Fukunaga K, Hirota S, et al: The expression profile of functional regulatory T cells, CD4⁺CD25^high+/forkhead box protein P3⁺, in patients with ulcerative colitis during active and quiescent disease. Clin Exp Immunol. 156:320–327. 2009. View Article : Google Scholar : PubMed/NCBI
31	Flower RJ: Drugs which inhibit prostaglandin biosynthesis. Pharmacol Rev. 26:33–67. 1974.PubMed/NCBI
32	Kim DS, Kim SJ, Kim MC, Jeon YD, Um JY and Hong SH: The therapeutic effect of chelidonic acid on ulcerative colitis. Biol Pharm Bull. 35:666–671. 2012. View Article : Google Scholar : PubMed/NCBI
33	Talero E, Sánchez-Fidalgo S, Villegas I, de la Lastra CA, Illanes M and Motilva V: Role of different inflammatory and tumor biomarkers in the development of ulcerative colitis-associated carcinogenesis. Inflamm Bowel Dis. 17:696–710. 2011. View Article : Google Scholar
34	Giriş M, Depboylu B, Doğru-Abbasoğlu S, Erbil Y, Olgaç V, Aliş H, Aykaç-Toker G and Uysal M: Effect of taurine on oxidative stress and apoptosis-related protein expression in trinitrobenzene sulphonic acid-induced colitis. Clin Exp Immunol. 152:102–110. 2008. View Article : Google Scholar
35	Lee MJ, Lee JK, Choi JW, et al: Interleukin-6 induces S100A9 expression in colonic epithelial cells through STAT3 activation in experimental ulcerative colitis. PLoS One. 7:e388012012. View Article : Google Scholar : PubMed/NCBI
36	Qiu W, Wu B, Wang X, Buchanan ME, Regueiro MD, Hartman DJ, Schoen RE, Yu J and Zhang L: PUMA-mediated intestinal epithelial apoptosis contributes to ulcerative colitis in humans and mice. J Clin Invest. 121:1722–1732. 2011. View Article : Google Scholar : PubMed/NCBI
37	Negoescu A, Guillermet C, Lorimier P, Brambilla E and Labat-Moleur F: Importance of DNA fragmentation in apoptosis with regard to TUNEL specificity. Biomed Pharmacother. 52:252–258. 1998. View Article : Google Scholar : PubMed/NCBI
38	Vermes I, Haanen C, Steffens-Nakken H and Reutelingsperger C: A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods. 184:39–51. 1995. View Article : Google Scholar : PubMed/NCBI
39	Samarghandian S, Afshari JT and Davoodi S: Honey induces apoptosis in renal cell carcinoma. Pharmacogn Mag. 7:46–52. 2011. View Article : Google Scholar : PubMed/NCBI
40	Lecoeur H: Nuclear apoptosis detection by flow cytometry: Influence of endogenous endonucleases. Exp Cell Res. 277:1–14. 2002. View Article : Google Scholar : PubMed/NCBI
41	Yu JZ, Wu XL, Yu B, Dai CQ, Liu KL, Guo-Qiang Q, Zhou GX, Lu SH, Ju H and Chen XY: The suppression effects of desacetyluvaricin on hepatocellular carcinoma and its possible mechanism. Pharmacogn Mag. 8:225–230. 2012. View Article : Google Scholar : PubMed/NCBI
42	Walters J, Pop C, Scott FL, Drag M, Swartz P, Mattos C, Salvesen GS and Clark AC: A constitutively active and uninhibitable caspase-3 zymogen efficiently induces apoptosis. Biochem J. 424:335–345. 2009. View Article : Google Scholar : PubMed/NCBI