醛固酮诱导人肾小管上皮细胞胶原Ⅰ、Ⅲ表达的研究  

Study on expressions of collagen Ⅰ and Ⅱ in aldosterone-induced human kidney tubular epithelial cells

作　　者：黄海燕[1] 魏佳莉[2] 

机构地区：[1]海南省人民医院中心实验室,海口570100 [2]海南省人民医院肾内科,海口570100

出　　处：《重庆医学》2016年第21期2900-2902,共3页Chongqing medicine

基　　金：海南省社会发展科技专项项目(2012SF04;2013SF04和2015SF43)

摘　　要：目的探讨Rho激酶对醛固酮(ALD)诱导的人肾小管上皮细胞胶原Ⅰ、Ⅲ(COLⅠ、Ⅲ)表达的影响。方法将人肾小管上皮细胞HK-2培养于含15%胎牛血清(FBS)的RPMI-1640培养液中,ALD受体拮抗剂依普利酮(10μmol/L)和Rho激酶抑制剂Y27632(1μmol/L)预处理细胞30min后,100nmol/L ALD作用HK-2细胞24h,实时定量PCR法检测各组细胞中COLⅠ、ⅢmRNA的表达,酶联免疫吸附试验测定培养上清液中COLⅠ、Ⅲ及Rho激酶蛋白表达水平。结果 ALD可上调HK-2细胞中COLⅠ、ⅢmRNA的表达,并增加培养上清液中Rho激酶、COLⅠ、Ⅲ蛋白表达水平,Rho激酶抑制剂Y27632及ALD受体拮抗剂依普利酮可拮抗上述效应。结论 ALD可活化HK-2细胞Rho激酶信号传导通路,并通过Rho激酶诱导HK-2细胞COLⅠ、Ⅲ的表达而加速肾小管间质纤维化的进展。Objective To investigate the role of Rho kinase on collagen（COL）Ⅰ and Ⅲ expression of human renal tubular epithelial cells.Methods Human kidney tubular epithelial（HK-2）cells were cultured in the RPMI-1640 culture solution containing15% fetal bovine serum.After 30 min pretreatment by the ALD receptor antagonist eplerenone（10μmol/L）and Rho kinase inhibitor Y27632（1μmol/L）,100nmol/L ALD acted the HK-2cells for 24 h.The expressions of collagenⅠandⅢ mRNA in each group were detected by real time PCR and the expression levels of COL Ⅰ,Ⅲ and Rho kinase protein were detected by ELISA.ResultsALD could up-regulate the expressions of COLⅠ,Ⅲ mRNA in HK-2cells,and increased the levels of Rho kinase,COLⅠ and Ⅲprotein,while the Rho kinase inhibitor Y27632 and ALD receptor inhibitor eplerenone could antagonize these effects.ConclusionALD could activate Rho kinase signal transduction pathway in HK-2cells and accelerate the progression of tubular interstitial fibrosis via Rho kinase induced expression of COL Ⅰ and Ⅲ.

关 键 词：醛固酮 胶原Ⅰ 胶原Ⅲ RHO激酶 人近端肾小管上皮细胞 肾小管间质纤维化 

分 类 号：R692[医药卫生—泌尿科学]