雷公藤多苷片对溃疡性结肠炎大鼠miR-146a、miR-146b及TLR4/MyD88依赖信号通路的调控作用研究  

Regulation of Tripterygium wilfordii Polycoride Tablet towards miR-146a, miR-146b, and TLR4/MyD88 dependent signaling pathway in ulcerative colitis rat model

作　　者：周毅骏 钦丹萍[2,1] 杨新艳[1] 杨雪静[3] 张春丽[4] 曹俊敏[3] 李艳平[3] 李珊[1] 

机构地区：[1]浙江中医药大学第一临床医学院,浙江杭州310053 [2]浙江中医药大学附属第一医院消化内科,浙江杭州310006 [3]浙江中医药大学附属第一医院中心实验室,浙江杭州310006 [4]浙江中医药大学附属第一医院病理科,浙江杭州310006

出　　处：《中草药》2016年第10期1723-1730,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81273903)

摘　　要：目的研究雷公藤多苷片(TWPT)对2,4,6-三硝基苯磺酸(TNBS)/乙醇溃疡性结肠炎(UC)大鼠微小RNA(mi R-146a、mi R-146b)及TLR4/My D88依赖信号通路的调控作用。方法采用TNBS/乙醇灌肠法制备UC大鼠模型。将90只雄性Wistar大鼠随机分为对照组,模型组,TWPT低、中、高剂量(30、60、120 mg/kg)组,硫唑嘌呤(AZA,60 mg/kg)组,每组15只。各组分别ig给予相应药物连续14 d。进行各组大鼠结肠组织大体及镜下病理评分。采用q RT-PCR法检测mi R-146a和mi R-146b的表达情况;Western blotting法和RT-PCR法检测大鼠结肠组织中TLR4/My D88依赖信号通路相关分子(TLR4、My D88、TRAF-6、NF-κB、TNF-α、IL-1β)在m RNA及蛋白水平的表达情况。结果 DAI评分,大体及镜下表现和评分均提示TNBS/乙醇UC大鼠模型造模成功,TWPT对UC大鼠临床症状的改善及黏膜愈合具有一定作用,该作用与AZA相比相当或强于AZA。q RT-PCR结果提示:与对照组相比,模型组中mi R-146a和mi R-146b表达明显增加(P<0.01);与模型组相比,TWPT及AZA均可显著抑制mi R-146a和mi R-146b的表达(P<0.01),其中TWPT中剂量组对2者的抑制作用最强。RT-PCR及Western blotting实验均提示:与对照组相比,模型组中TLR4/My D88依赖信号通路相关的分子无论在m RNA还是蛋白水平表达均显著升高(P<0.01);与模型组相比,TWPT呈剂量依赖性地抑制该信号通路上各节点分子m RNA及蛋白水平的表达,其中TWPT高剂量组中各节点分子m RNA及蛋白表达水平低于模型组(P<0.05)。与AZA组比较,TWPT高剂量组对该信号通路上游因子(TLR4、My D88、TRAF-6、NF-κB)m RNA及蛋白表达水平的抑制作用略好于AZA组,而对末端炎症因子TNF-α及IL-1βm RNA及蛋白水平的抑制作用却略逊于AZA组,但上述2种差异均无统计学意义(P>0.05)。结论在TNBS/乙醇UC大鼠模型中,TWPT能通过抑制mi R-146a和mi R-146b的表达,并能抑制TLR4/My D88依赖信号通路及炎症因子IL-1β及TNF-α释放,对信号Objective To study the regulatory effect of Tripterygium wilfordii Polycoride Tadlet（TWPT） towards mi R-146 a, mi R-146 b, and TLR4/My D88 dependent signaling pathway in TNBS/ethanol ulcerative colitis（UC） rat model. Methods TNBS enema was adopted to build TNBS/ethanol UC rat model. After the modeling procedure, 90 male Wistar rats were divided into six groups, including normal, model, low-, mid-, high-dose TWPT, and azathioprine（AZA） groups, and each for 15 rats. All rats in each group were administered with corresponding medicines for 14 d. After 14 d administration, corresponding colon tissues were taken to undergo general and microscopic evaluation. q PCR was adopted to test the expression of mi R-146 a and mi R-146 b. Western blotting analysis and RT-PCR were adopted to test the m RNA and protein expression levels of TLR4/My D88 dependent signaling pathway related molecular, including TLR4, My D88, TRAF-6, NF-κB, TNF-α, and IL-1β. Results DAI, general and microscopic evaluation all showed that TNBS/ethanol UC rat model was successfully established. TWPT could improve UC-related clinical manifestation and promote the colonic mucosa healing procedure and such effect was equal to AZA. q RT-PCR showed that the expression of mi R-146 a and mi R-146 b in model group was significantly superior to that in normal group（P〈0.01）. Compared with the model group, TWPT and AZA could significantly inhibit the expression of mi R-146 a and mi R-146b（P〈0.01）. The mid-dose TWPT showed the strongest inhibitory effect. RT-PCR and Western blotting results showed that the expression of TLR4/My D88 dependent signaling pathway related molecular in model group was significantly superior to that in normal group either in m RNA or protein levels（P〈0.01）. Compared with model group, TWPT could inhibit the expression of each spot in TLR4/My D88 dependent signaling pathway in a dose-dependent manner. The inhibitory effect of high-dose TWPT towards the above molecular was superior to that in model group eithe

关 键 词：雷公藤多苷片 溃疡性结肠炎 MIR-146A miR-146b TLR4/MyD88依赖信号通路 

分 类 号：R285.5[医药卫生—中药学]