白藜芦醇对小鼠溃疡性结肠炎的影响*

doi:10.12047/j.cjap.5826.2019.097

摘要/Abstract

摘要： 目的:研究白藜芦醇通过调节Wnt/β-catenin信号通路抗溃疡性结肠炎的作用机制。方法:①葡聚糖硫酸钠盐(DSS)诱发溃疡性结肠炎实验:28只C57BL/6小鼠随机分为4组(n=7):control组、 DSS组、DSS+白藜芦醇(DSS+Res)组和Res组。实验周期为3周,小鼠饮用DSS水诱导溃疡性结肠炎并给予白藜芦醇灌胃。实验期间每天称小鼠体重并观察小鼠活动和粪便情况。处理结束后,安乐死小鼠,取小鼠脾脏称重,取小鼠结肠测量长度。苏木精-伊红染色法(H&E)染色观察小鼠结肠组织病理改变;实时荧光定量PCR(qPCR)检测小鼠结肠组织miRNA-31的表达;Western Blot检测小鼠结肠组织β-catenin、Cyclin D1蛋白的表达。②离体实验:以10 mg/ml浓度的白藜芦醇处理HCT 116细胞,检测HCT 116细胞β-catenin、低密度脂蛋白受体相关蛋白6(LRP-6)、卷曲蛋白3(FZD3)、c-Myc蛋白的表达;HCT 116细胞转染miRNA-31 mimic和inhibitor,检测β-catenin蛋白的表达。结果:①DSS组小鼠实验期间体重下降明显,精神萎靡,活动减少,出现血便;处理结束后小鼠的结肠长度缩短,脾脏增大。而给予白藜芦醇后小鼠的以上情况得到改善。②白藜芦醇抑制了溃疡性结肠炎小鼠结肠组织miRNA-31的表达及β-catenin、Cyclin D1蛋白的表达。③白藜芦醇下调HCT 116细胞β-catenin、LRP-6、FZD3、c-Myc蛋白的表达。转染miRNA-31 inhibitor后,HCT 116细胞中β-catenin蛋白表达减少。结论:白藜芦醇能够抑制DSS诱导的小鼠溃疡性结肠炎,这种作用与下调Wnt信号通路有关,其对Wnt 信号的下调作用与miRNA-31有关。

关键词: 白藜芦醇, Wnt/β-catenin信号通路, miRNA-31, 溃疡性结肠炎, 小鼠

Abstract: Objective: To investigate the anti-ulcerative colitis mechanism of resveratrol through regulation of Wnt/β-catenin signaling pathway. Methods: ①The experiment of ulcerative colitis induced by dextran sulfate sodium salt (DSS): 28 C57BL/6 mice were randomly divided into four groups including control group(n=7), DSS group(n=7), DSS+Resveratrol (DSS+Res) group(n=7) and Res group(n=7). The experiment lasted for 3 weeks. Ulcerative colitis of mice was induced by drinking DSS water and treated with resveratrol by intragastric administration. The mice were weighed daily and their activities and state of feces were recorded. After that, the mice were euthanized, the spleens were weighed, and the colonic length was measured.Hematoxylin-eosin staining (HE) was used to observe the pathological changes of the colon, and the expression of miR-31 in colonic tissue was detected by quantitative real-time PCR (qPCR). The expressions of β-catenin and Cyclin D1 were measured by Western blot. ②In vitro experiment: HCT 116 cells were treated with resveratrol at 10 mg/ml, the expressions of β-catenin, LDL receptor related protein-6 (LRP-6), frizzled-3 (FZD3) and c-Myc were detected. The expression of β-catenin was also detected in HCT 116 cells transfected with miRNA-31 mimic and miRNA-31 inhibitor. Results: ① The body weight was decreased in DSS group, the activity was decreased and blood stool appeared. The colonic length of mice was shortened, the spleen was enlarged and the tissue was damaged seriously in DSS group. While the above symptoms were improved after resveratrol treatment. ② Resveratrol inhibited the expressions of miRNA-31, β-catenin and CyclinD1 in ulcerative colitis mice, and also down-regulated the expressions of β-catenin, LRP-6, FZD3 and c-Myc in HCT 116 cells. After transfection of miRNA-31 inhibitor, the expression of β-catenin was decreased in HCT 116 cells. Conclusion: Resveratrol suppresses DSS induced colitis by down-regulation of Wnt signal pathway. The down-regulation of Wnt signal may be related to miRNA-31.

Key words: resveratrol, Wnt/ β-catenin signaling pathway, MiRNA-31, ulcerative colitis, mice

刘鑫, 吴亚俐, 刘凯丽, 崔香丽, 都新新, 张文琴. 白藜芦醇对小鼠溃疡性结肠炎的影响^*[J]. 中国应用生理学杂志, 2019, 35(5): 447-453.

LIU Xin, WU Ya-li, LIU Kai-li, CUI Xiang-li, DU Xin-xin, ZHANG Wen-qin. Effects of resveratrol on ulcerative colitis in mice and its mechanism[J]. CJAP, 2019, 35(5): 447-453.

参考文献

[1] Ungaro R, Mehandru S, Allen PB, et al. Ulcerative colitis[J]. Lancet, 2017, 389(10080): 1756-1770.
[2] Feuerstein JD, Cheifetz AS. Ulcerative colitis: epidemiology, diagnosis, and management[J]. Mayo Clin Proc, 2014, 89(11): 1553-1563.
[3] Jess T, Rungoe C, Peyrin-Biroulet L. Risk of colorectal cancer in patients with ulcerative colitis: a meta-analysis of population-based cohort studies[J]. Clin Gastroenterol Hepatol, 2012, 10(6): 639-645.
[4] Cosin-Roger J, Ortiz-Masia D, Calatayud S, et al. M2 macrophages activate WNT signaling pathway in epithelial cells: relevance in ulcerative colitis[J]. PLoS One, 2013, 8(10): e78128.
[5] Shenoy AK, Fisher RC, Butterworth EA, et al. Transition from colitis to cancer: high Wnt activity sustains the tumor-initiating potential of colon cancer stem cell precursors[J]. Cancer Res, 2012, 72(19): 5091-5100.
[6] Shimodate Y, Takanashi K, Waga E, et al. Exacerbation of bloody diarrhea as a side effect of mesalamine treatment of active ulcerative colitis[J]. Case Rep Gastroenterol, 2011, 5(1): 159-165.
[7] Marteau P, Nelet F, Lelu M, et al. Adverse events in patients treated with 5-aminosalicylic acid: 1993-1994 pharmacovigilance report for Pentasa in France[J]. Aliment Pharmacol Ther, 1996, 10(6): 949-956.
[8] Fiorentini MT, Fracchia M, Galatola G, et al. Acute pancreatitis during oral 5-aminosalicylic acid therapy[J]. Dig Dis Sci, 1990, 35(9): 1180-1182.
[9] Bressler B, Marshall JK, Bernstein CN, et al. Clinical practice guidelines for the medical management of nonhospitalized ulcerative colitis: the Toronto consensus[J]. Gastroenterology, 2015, 148(5): 1035-1058.
[10]Hoentjen F1, Sakuraba A, Hanauer S. Update on the management of ulcerative colitis[J]. Curr Gastroenterol Rep, 2011, 13(5): 475-485.
[11]Ji Q, Liu X, Fu X, et al. Resveratrol inhibits invasion and metastasis of colorectal cancer cells via MALAT1 mediated Wnt/beta-catenin signal pathway[J]. PLoS One, 2013, 8(11): e78700.
[12]Yao J, Wei C, Wang JY, et al. Effect of resveratrol on Treg/Th17 signaling and ulcerative colitis treatment in mice[J]. World J Gastroenterol, 2015, 21(21): 6572-6581.
[13]Samsamikor M, Daryani NE, Asl PR, et al. Resveratrol supplementation and oxidative/anti-oxidative status in patients with ulcerative colitis: a randomized, double-blind, placebo-controlled pilot study[J]. Arch Med Res, 2016, 47(4): 304-309.
[14]Yuan SX, Wang DX, Wu QX, et al. BMP9/p38 MAPK is essential for the antiproliferative effect of resveratrol on human colon cancer[J]. Oncol Rep, 2016, 35(2): 939-947.
[15]Li D, Wang G, Jin G, et al. Resveratrol suppresses colon cancer growth by targeting the AKT/STAT3 signaling pathway[J]. Int J Mol Med, 2019, 43(1): 630-640.
[16]Klein U, Lia M, Crespo M, et al. The DLEU2/miR-15a/16-1 cluster controls B cell proliferation and its deletion leads to chronic lymphocytic leukemia[J]. Cancer Cell, 2010, 17(1): 28-40.
[17]Bonci D, Coppola V, Musumeci M, et al. The miR-15a-miR-16-1 cluster controls prostate cancer by targeting multiple oncogenic activities[J]. Nat Med, 2008, 14(11): 1271-1277.
[18]王茜, 崔香丽. 白藜芦醇通过下调miR-31-5p的表达抑制细胞增殖[D]. 山西医科大学, 2018: 1～80
[19]宋娟, 王佳, 李宝红, 等. PI3K/Akt信号通路在白藜芦醇抗大鼠缺血/再灌注性心律失常中的作用及机制[J]. 中国应用生理学杂志, 2017, 33(3) : 239-243.
[20]Samsami-Kor M, Daryani NE, Asl PR, et al. Anti-inflammatory effects of resveratrol in patients with ulcerative colitis: A randomized, double-blind, placebo-controlled pilot study[J]. Arch Med Res, 2015, 46(4): 280-285.
[21]Yao J, Wang J Y, Liu L, et al. Anti-oxidant effects of resveratrol on mice with DSS-induced ulcerative colitis[J]. Arch Med Res, 2010, 41(4): 288-294.
[22]Cui X, Jin Y, Hofseth AB, et al. Resveratrol suppresses colitis and colon cancer associated with colitis[J]. Cancer Prev Res (Phila), 2010, 3(4): 549-559.
[23]Abdallah DM, Ismael NR. Resveratrol abrogates adhesion molecules and protects against TNBS-induced ulcerative colitis in rats[J]. Can J Physiol Pharmacol, 2011, 89(11): 811-818.
[24]Macdonald BT, Tamai K, He X. Wnt/beta-catenin signaling: components, mechanisms, and diseases[J]. Dev Cell, 2009, 17(1): 9-26.
[25]Clevers H, Loh KM, Nusse R. Stem cell signaling. An integral program for tissue renewal and regeneration: Wnt signaling and stem cell control[J]. Science, 2014, 346(6205): 1248012.
[26]Rhodes JM, Campbell BJ. Inflammation and colorectal cancer: IBD-associated and sporadic cancer compared[J]. Trends Mol Med, 2002, 8(1): 10-16.
[27]Koch S. Extrinsic control of Wnt signaling in the intestine[J]. Differentiation, 2017, 97: 1-8.
[28]黄循铷, 王承党, 王瑞幸, 等. 溃疡性结肠炎小鼠肠道通透性改变与TNF-α及NF-κB P65的关系[J]. 中国应用生理学杂志, 2016, 32(2) : 112-115.
[29]Goretsky T, Bradford EM, Ryu H, et al. A cytosolic multiprotein complex containing p85alpha is required for beta-catenin activation in colitis and colitis-associated cancer[J]. J Biol Chem, 2016, 291(8): 4166-4177.
[30]Cosin-Roger J, Ortiz-Masia D, Calatayud S, et al. The activation of Wnt signaling by a STAT6-dependent macrophage phenotype promotes mucosal repair in murine IBD[J]. Mucosal Immunol, 2016, 9(4): 986-998.
[31]Clevers H, Nusse R. Wnt/beta-catenin signaling and disease[J]. Cell, 2012, 149(6): 1192-1205.
[32]Wang CJ, Stratmann J, Zhou ZG, et al. Suppression of microRNA-31 increases sensitivity to 5-FU at an early stage, and affects cell migration and invasion in HCT-116 colon cancer cells[J]. BMC Cancer, 2010, 10: 616.
[33]Chen T, Yao L Q, Shi Q, et al. MicroRNA-31 contributes to colorectal cancer development by targeting factor inhibiting HIF-1alpha (FIH-1)[J]. Cancer Biol Ther, 2014, 15(5): 516-523.

白藜芦醇对小鼠溃疡性结肠炎的影响^*

Effects of resveratrol on ulcerative colitis in mice and its mechanism

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