贝那普利对高糖培养下大鼠肾小球系膜细胞整合素连接激酶及α平滑肌肌动蛋白表达的影响  

Effects of benazepril on the expressions of intergrin-linked kinase and eL-smooth muscle actin in glomerular mesangial cells exposed to high glucose

作　　者：牛洪琳[1] 李英[1] 刘茂东[1] 迟雁青[1] 张涛[1] 王秀芬[1] 

机构地区：[1]河北医科大学第三医院肾内科,石家庄050081

出　　处：《中华肾脏病杂志》2013年第1期33-38,共6页Chinese Journal of Nephrology

摘　　要：目的观察贝那普利对高糖培养下肾小球系膜细胞（GMC）中整合素连接激酶（ILK）及α平滑肌肌动蛋白（α-SMA）表达的影响，探讨贝那普利肾脏保护作用的调节机制。方法将GMC常规培养分为4组：正常糖对照组（D-葡萄糖5．5mmol／L，NG组）、甘露醇组（甘露醇20mmo／／L，MG组）、高糖组（D-葡萄糖30mmol／L，HG组）、高糖＋贝那普利干预组（D-葡萄糖30mmol／L＋贝那普利10μmol／L，ACEI组）。于实验开始后3h、6h、12h、24h、48h、72h分别收获4组细胞，应用RT—PCR法、Western印迹法和细胞免疫荧光法检测各组GMC中ILK、α-SMA的表达。结果高糖环境下GMC中ILK、α-SMAmRNA和蛋白表达水平较正常对照组显著升高（均P〈0．05），ILK在48h达到最高（P〈0．05），α-SMA在72h达到最高（P〈0．01）。贝那普利干预后上述指标较高糖组表达水平下降，但未能恢复至正常糖对照组水平。同时间点甘露醇组ILKmRNA和蛋白水平与正常糖组差异无统计学意义（P〉0．05）；同时间点甘露醇组α-SMAmRNA和蛋白水平显著高于正常糖对照组（均P〈0．05）。结论贝那普利可能通过降低ILK和α-SMA的异常表达来延缓糖尿病肾病的肾脏纤维化进程和高糖介导的系膜细胞表型转化。系膜细胞在糖尿病。肾病中的表型转化可能同时存在渗透浓度依赖性。Objective To investigate the effect of benazepril on intergrin-linked kinase （ILK） and α-smooth muscle actin （α-SMA） expression in glomerular mesangial cells induced by high-glucose. Methods The mesangial cells from SD rat （HBZY- 1） were cultured conventionally and randomly divided into four groups： normal glucose （D-glucose 5.5 mmol/L, group NG）, mannitol-treated group （mannitol 20 mmol/L, group MG）, high glucose （D-glucose 30 mmol/L, group HG）, Benazepril-treated high glucose group （D-glucose 30 mmol/L ＋ Benazepril 10 μmol/L, group ACEI）. Cells from NG, MG, HG, ACEI gronps were harvested after 3, 6, 12, 24, 48 and 72 hours of treatment respectively. The mRNA expressions of ILK and α- SMA were detected by RT- PCR. The protein levels of ILK and α- SMA were detected by Western blotting and immunofluorescence. Results The expressions of ILK mRNA and protein in HG group were significantly increased compared with those in NG group （all P 〈 0.05）. The increased expressions of ILK and α- SMA in HG group were time- dependent and theexpression reached the peak at 48 h （ILK, P 〈 0.05） or 72 h （α-SMA, P 〈 0.01）. The expressions of ILK and α-SMA in ACEI group were lower than those in HG group （all P 〈 0.01）, but failed to rescue to the same level as those in NG. There was no significant differences of ILK expressions between MG group and NG group at the same time point （P 〉 0.05）. The expressions of α- SMA mRNA and protein in MG were higher than that in NG （P 〈 0.05）, which suggest that high osmotic pressure could cause the increasing of α-SMA. Conclusions Benazepril can decrease the expressions of ILK and α-SMA to inhibit the process of fibrosis in DN and mediate the phenotypic transformation of glomerular mesangial cells. The phenotypic transformation of glomerular mesangial ceils in glucose may also depend on high osmotic pressure in DN.

关 键 词：糖尿病肾病 系膜细胞 纤维化 整合素连接激酶 贝那普利 

分 类 号：R692[医药卫生—泌尿科学] R587[医药卫生—外科学]