地马煎剂对溃疡性结肠炎大鼠肠上皮细胞BNIP3蛋白与mRNA的影响  

Effect of Dima Decoction on BNIP3 Protein and mRNA Expressions in Intestinal Epithelial Cells of Ulcerative Colitis Mice

作　　者：李一芳[1] 王睿[2] 查安生[2] 王丽娜[1] 谭辉[1] LI Yi-fang;WANG Rui;ZHA An-sheng;WANG Li-na;TAN Hui(Anhui University of Chinese Medicine,Hefei 230012,China;First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学,合肥230012 [2]安徽中医药大学第一附属医院,合肥230031

出　　处：《中国实验方剂学杂志》2020年第11期107-112,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：安徽省自然科学基金青年基金项目(1808085QH257);安徽省卫生计生委中医药科研课题项目(2016zy68)。

摘　　要：目的:通过观察地马煎剂对腺病毒E1B 19 kD相互作用蛋白-3(BNIP3)蛋白及mRNA的调控水平,了解其在干预溃疡性结肠炎(UC)线粒体自噬及控制炎症中的机制。方法:60只SPF级SD大鼠随机分为空白组,模型组,地马煎剂低、中、高剂量(8.5,17.0,34.0 g·kg^-1)组,美沙拉嗪(3.0 g·kg^-1)组,每组10只。模型组高脂饮食+2,4,6-三硝基苯磺酸(TNBS)+乙醇法建立活动期UC大鼠模型,给药组连续灌胃14 d。苏木素-伊红(HE)染色观察大鼠结肠黏膜组织病理变化,蛋白免疫印迹法(Western blot)及实时荧光定量聚合酶链式反应(Real-time PCR)检测肠上皮细胞BNIP3蛋白及mRNA的表达。结果:HE染色结果显示,模型组大鼠结肠组织可见大量炎性细胞浸润以及非干酪样肉芽组织,而各给药组大鼠出现不同程度的纤维性修复等表现。与正常组比较,模型组BNIP3蛋白及mRNA表达均明显升高(P<0.05);与模型组比较,地马煎剂低、中、高剂量组及美沙拉嗪组BNIP3蛋白及mRNA表达均显著升高(P<0.01);与地马煎剂低剂量组比较,地马煎剂中、高剂量组及美沙拉嗪组BNIP3蛋白及mRNA表达明显升高(P<0.05)。结论:地马煎剂可增加UC活动期大鼠肠上皮的BNIP3表达,这可能是其促进线粒体自噬对抗UC炎症的机制之一。Objective:To study the regulatory level of Dima decoction on adenovirus E1B 19 kD interacting protein 3(BNIP3)protein and to understand the mechanism in intervening mitophagy and controlling inflammation in ulcerative colitis.Method:The 60 SPF SD rats were randomly divided into normal control group,model control group,low,medium ang high dose Dima decoction(8.5,17.0,34.0 g·kg^-1)group and mesalazine(3.0 g·kg^-1)group,10 for each group.The model of UC rats was established by the method of environmental diet intervention+2,4,6-trinitro-Benzenesulfonicacid(TNBS)+ethanol,the administration group was administered for 14 days.Hematoxylin-eosin(HE)was used to detect pathological changes of colonic mucosal tissues in rats.Western blot and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)were used to detect the expressions of BNIP3 protein and mRNA in intestinal epithelial cells.Result:HE staining results showed that a large number of inflammatory cell infiltration and non-cheesy granulation tissue were observed in the colon of rats in the model group,while the rats in each drug group showed different degrees of fibrous repair.Compared with normal group,the expression of BNIP3 protein and mRNA in model group were significantly increased(P<0.05).Compared with model group,the expression of BNIP3 protein and mRNA in low,medium and high-dose Dima decoction group and mesalazine group were significantly increased(P<0.01).Compared with low-dose group,the expression of BNIP3 protein and mRNA in medium and high-dose groups and mesalazine group were significantly increased(P<0.05).Conclusion:Dima decoction can increase the expression of BNIP3 in the intestinal epithelium of UC active rats,which may be one of the mechanisms of promoting mitochondrial autophagy against UC inflammation.

关 键 词：地马煎剂 溃疡性结肠炎 腺病毒E1B 19kD相互作用蛋白-3(BNIP3) 线粒体自噬 机制 

分 类 号：R2-0[医药卫生—中医学] R289