法舒地尔对高糖培养肾小管上皮细胞增殖和纤维化的影响  

Influence of fasudil on the proliferation and fibrosis of renal tubular epithelial cells incubated with high glucose

作　　者：顾玲佳[1] 倪连松[2] 

机构地区：[1]台州市第一人民医院内分泌科,浙江台州318020 [2]温州医科大学附属第一医院内分泌科,浙江温州325015

出　　处：《温州医学院学报》2014年第10期723-726,732,共5页Journal of Wenzhou Medical College

基　　金：温州市科技局科研基金资助项目(Y20100020)

摘　　要：目的:探讨法舒地尔(Fasudil)对高糖培养的肾小管上皮细胞(HK-2)增殖和纤维化的影响。方法:将HK-2细胞分为正常对照组(NG组,葡萄糖浓度5.5 mmol/L)、高张组(HM组,葡萄糖浓度5.5 mmol/L+甘露醇54.5 mmol/L)、高糖组(HG组,葡萄糖浓度60 mmol/L)、高糖+小剂量法舒地尔干预组[FA(5)组,D-葡萄糖60 mmol/L+法舒地尔5μmol/L]、高糖+中剂量法舒地尔干预组[FA(10)组,D-葡萄糖60 mmol/L+法舒地尔10μmol/L];6高糖+大剂量法舒地尔干预组[FA(20)组,D-葡萄糖60 mmol/L+法舒地尔20μmol/L]。四甲基偶氮唑盐(MTT)法测定HK-2细胞增殖;免疫共沉淀法检测p-MYPT1-Thr853;Western印迹法检测结缔组织生长因子(CTGF)的表达;ELISA法检测细胞培养上清液转化生长因子β1(TGF-β1)和纤维连接蛋白(Fn)的含量。结果:与NG组比,HG组HK-2细胞出现增殖增加,p-MYPT1-Thr853、TGF-β1、CTGF和Fn的表达增加。与HG组比,FA(5)组、FA(10)组、FA(20)组均抑制了HK-2细胞的过度增殖,p-MYPT1-Thr853、TGF-β1、CTGF和Fn的表达均减少。结论:法舒地尔可能通过抑制Rho激酶活性抑制高糖培养的肾小管上皮细胞增殖和纤维化。Objective： To investigate the influence of fasudil on the proliferation and fibrosis of human renal tubular epithelial cells incubated with high glucose. Methods： HK-2 cells were divided into normal control group （D-glucose 5.5 mmol/L）, high tension group （D-glucose 5.5 mmol/L＋54.5 mmol/L mannitol）, high glucose group （D-glucose 60 mmol/L）, high glucose＋fasudil treatment group （the concentrations of fasudil were 5, 10 and 20 μmol/L）. Cell proliferation was assessed with MTT assay. Changes in the p-MYPT1-Thr853 were detected with co-immunoprecipitation assay. The expression of connective tissue growth factor （CTGF） was detected with western blot, the protein content of transforming growth factor-β1 （TGF-β1） and fibronectin （Fn） in the superna- tant were detected by ELISA. Results： Compared with normal control group, cell proliferation and the expres- sion of p-MYPT1-Thr853, TGF-β1, CTGF and Fn were significantly increased in high glucose group. Compared with high glucose group, cell proliferation and the expression of p-MYPT1-Thr853, TGF-β1, CTGF and Fn were significantly decreased in fasudil treatment group. Conclusion： Fasudil can inhibit high glucose-induced prolif- eration and fibrosis of HK-2 and it may produce effect through the inhibition ofRho kinase activity.

关 键 词：法舒地尔 肾小管上皮细胞 纤维化 糖尿病肾病 

分 类 号：R965.2[医药卫生—药理学]