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（COL Ⅰ、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30 min后，100 nmol／L ALD作用 HK‐2细胞24 h ，实时定量 PCR法检测各组细胞中COLⅠ、Ⅲ mRNA的表达，酶联免疫吸附试验测定培养上清液中COL Ⅰ、Ⅲ及Rho激酶蛋白表达水平。结果 ALD可上调HK‐2细胞中COL Ⅰ、Ⅲ mRNA的表达，并增加培养上清液中Rho激酶、COL Ⅰ、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COL Ⅰ、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 containing 15% fetal bovine serum .After 30 min pretreatment by the ALD receptor antagonist eplerenone(10 μmol/L) and Rho kinase inhibi‐tor Y27632(1 μmol/L) ,100 nmol/L ALD acted the HK‐2 cells for 24 h .The expressions of collagenⅠ and Ⅲ 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 .Results ALD could up‐regulate the expressions of COLⅠ ,Ⅲ mRNA in HK‐2 cells ,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 .Conclusion ALD could activate Rho kinase signal transduction pathway in HK‐2 cells and accelerate the progression of tubular interstitial fibro‐sis via Rho kinase induced expression of COL Ⅰ and Ⅲ .
展开▼
