乌梅丸通过调控miR-146a对溃疡性结肠炎大鼠结肠细胞凋亡的影响及机制研究

Study on effect of Wumei Pill on apoptosis of colon cells in rats with ulcerative colitis and its mechanism by regulating miR-146a

目的:探讨乌梅丸通过调控miR-146a对溃疡性结肠炎(UC)大鼠结肠细胞凋亡的影响及机制。方法:选取50只SD大鼠,每组10只,分为正常对照组、模型组、乌梅丸低、中、高剂量组,转染anti-miR-NC、anti-miR-146a、miR-NC、miR-146a,作为anti-miR-NC组、anti-miR-146a组、乌梅丸+miR-NC组、乌梅丸+miR-146a组。CCK-8检测细胞增殖;流式细胞术检测细胞凋亡;RT-qPCR检测细胞miR-146a表达;ELISA检测细胞IL-1β、IL-13含量;Western blot检测细胞B细胞淋巴瘤2(Bcl-2)、Bax蛋白表达。结果:与正常对照组比较,模型组细胞存活率、Bcl-2蛋白表达降低,细胞凋亡率、Bax蛋白表达、IL-1β、IL-13含量、miR-146a表达升高(P<0.05);与模型组比较,乌梅丸显著升高细胞存活率、Bcl-2蛋白表达,降低细胞凋亡率、Bax蛋白表达、IL-1β、IL-13含量、miR-146a表达(P<0.05)。抑制miR-146a可提高细胞存活率、Bcl-2蛋白表达,降低细胞凋亡率、IL-1β、IL-13含量、Bax蛋白表达(P<0.05)。过表达miR-146a表达逆转乌梅丸对UC大鼠结肠细胞增殖和凋亡的影响。结论:乌梅丸通过上调miR-146a表达降低UC大鼠结肠细胞凋亡。

Objective:To investigate effect of Wumei Pill on colon cell apoptosis in ulcerative colitis(UC)rats by regulating miR-146a and its mechanism.Methods:A total of 50 SD rats were selected,with 10 rats in each group,and divided into control group,model group,Wumei Pill low,medium and high doses groups.Transfected with anti-miR-NC,anti-miR-146a,miR-NC,miR-146a as anti-miR-NC group,anti-miR-146a group,Wumei Pill+miR-NC group,Wumei Pill+miR-146a group.CCK-8 was used to de-tect cell proliferation;flow cytometry was used to detect cell apoptosis;RT-qPCR was used to detect cell miR-146a expression;ELI-SA was used to detect cell IL-1βand IL-13 contents;Western blot was used to detect expressions of B-cell lymphoma 2(Bcl-2)and Bax proteins.Results:Compared with control group,cell survival rate,Bcl-2 protein expression in model group were decreased,while apoptosis rate,Bax protein expression,IL-1β,IL-13 contents and miR-146a expression were increased(P<0.05).Compared with model group,Wumei Pill significantly increased cell survival rate and Bcl-2 protein expression,decreased cell apoptosis rate,Bax protein expression,IL-1β,IL-13 contents and miR-146a expression(P<0.05).Inhibition of miR-146a increased cell survival rate,Bcl-2 protein expression,decreased cell apoptosis rate,IL-1β,IL-13 contents and Bax protein expression(P<0.05).Overexpression of miR-146a reversed effects of Wumei Pills on proliferation and apoptosis of colon cells in UC rats.Conclusion:Wumei Pill can reduce apoptosis of colon cells in UC rats by up-regulating expression of miR-146a.