糖肾平含药血清对高糖诱导肾小管上皮细胞增殖作用及RhoA/ROCK信号通路影响的研究  

Effect of Tangshenping Medicated Serum on Proliferationand RhoA/ROCK Signaling Pathway of High Glucose-induced Epithelial Cells of Renal Tubules

作　　者：苗永辉[1] 赵宗江[1] 杨冠男[1] 王婷[1] 黄雅薇 张紫嫣 

机构地区：[1]北京中医药大学中医学院,北京100029

出　　处：《世界科学技术-中医药现代化》2017年第6期1050-1056,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然基金委面上项目(81373831):基于"肾痿"组方的糖肾平干预糖尿病肾病肾小管上皮细胞转分化的分子机制研究;负责人:赵宗江

摘　　要：目的:探讨糖肾平含药血清对高糖刺激肾小管上皮细胞增殖作用及Rho A/ROCK信号通路的影响。方法:制备鼠含药血清;培养肾小管上皮细胞分为:正常组,高糖组,抑制剂(Y27632)组,厄贝沙坦组,糖肾平低、中、高剂量组,3000细胞/孔种于96孔板,每组8个复孔,12、24、48、60 h观察细胞,MTT法检测细胞增殖。Western blotting检测肾小管上皮细胞中Rho A、ROCK1、α-SMA和E-Cadherin蛋白的表达。结果:高糖诱导后细胞呈梭型或不规则三角形;含药血清干预后,细胞呈扁平不规则多角形。MTT:12 h,与正常组比,各组OD值升高;24、48、60 h,与正常组比,高糖组OD值显著升高(P<0.01);与高糖组比,各治疗组OD值有不同程度降低(P<0.05),48 h,与Y27632组、厄贝沙坦组和糖肾平高剂量组比,糖肾平低、中剂量组OD值降低(P<0.05);60 h,与Y27632组比,糖肾平中剂量组降低;与厄贝沙坦组比,糖肾平各剂量组OD值降低(P<0.05);与糖肾平高剂量组比,糖肾平低剂量组升高(P<0.05)。Western blotting,与正常组比,高糖组E-Cadherin蛋白表达减少,Rho A、ROCK1和α-SMA蛋白表达增加(P<0.01);与高糖组比,各组E-Cadherin蛋白表达增加,Rho A、ROCK1和α-SMA蛋白表达减少,高剂量组有显著差异(P<0.01);与Y27632组比,糖肾平高剂量组ROCK1、E-Cadherin和α-SMA蛋白表达无统计学差异(P>0.05),Rho A蛋白表达减少(P<0.01);与厄贝沙坦组比,糖肾平高剂量组Rho A、ROCK1、E-Cadherin和α-SMA蛋白表达无统计学差异(P>0.05)。结论:糖肾平通过抑制高糖诱导的肾小管上皮细胞增殖和Rho A/ROCK信号通路相关分子的表达,从而逆转上皮-间质转分化,抑制肾间质纤维化,延缓糖尿病肾病病情进展。To study the effect of the Tangshenping containing serum on the proliferation of the high glucose-induced epithelial cells of renal tubules. To prepare durg-contained serum from rats to enter into in vitor reaction system, the cellscultured via 10% FBS-RPMI 1640 were randomly divided into 7groups：the normal group, themodel group, the Y27632 group, theirbesartangroup, the small dose Tangshenping group, the medium dose Tangshenping group and the high dose Tangshenping group and The cells were cultured in 3000 cells/well and grown in 96-well plates. Each group had 8 wells, then detect the effects of all serum sections on the proliferation of high glucose-induced epithelial cells of kidney tubules by the MTT colorimetric method after cultured for 12 h, 24 h, 48 h, and 60 h. Based on the resuhs above, cell protein were extracted from each group at 24 h, and the expression of RhoA, ROCK1, α-SMA and E-cadherin in each group were detected by Western blotting. After high glucose stimulation, the shape of cell was shuttle-like or irregular triangle, the way it grew was radial; after the intervention of the corresponding serum, the shape of the cell was flat and irregular polygonal.Started with 12h,compared with the normal group, OD value of other groups increased; at the 24h, 48hand 60h, compared with the normal group, OD value of high glucose groupincreased significantly （P〈0.01）; compared with the high glucose group, OD value of treatment groups decreased （P〈0.05）; and 48 h, compared with the Y27632group,irbesartan groupand Tangshenping high dose group, OD value of Tangshenping low and medium dose groups decreased （P〈0.05）; 60 h, compared with Y27632 group, OD value of Tangshenping medium dose groups decreased;compared with irbesartan group, OD value of o Tangshenpinggroupsdecreased （P〈0.05）; compared with Tangshenping high dose group, OD value of Tangshenpinglow groupsdecreased （P〈0.05） Western blotting analysis showed that compared with normal group, the expression of E-Cadherin protein

关 键 词：糖肾平 肾痿 肾小管上皮细胞株 KKAY小鼠 细胞增殖 MTT EMT RhoA/ROCK信号通路 

分 类 号：R285.5[医药卫生—中药学]