法舒地尔对高糖培养的肾小管上皮细胞转分化的抑制作用  

Inhibitory effect of fasudil on transdifferentiation of renal tubular epithelial cells incubated with high concentration of glucose

作　　者：倪连松[1] 高倩[1] 顾玲佳[1] 

机构地区：[1]温州医学院附属第一医院内分泌科,浙江温州325000

出　　处：《中国药理学与毒理学杂志》2012年第1期30-34,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：温州市科技局基金资助(Y20100020)~~

摘　　要：目的探讨法舒地尔对高糖培养人肾小管上皮细胞(HK-2)转分化的影响及可能作用机制。方法人肾小管上皮细胞同时加入葡萄糖60 mmol·L-1和法舒地尔20μmol·L-1。配体结合沉淀法检测葡萄糖60mmol·L-1作用0～24 h后细胞Rho A活性,免疫细胞化学技术检测上皮钙黏素表达,Western印迹法检测α-平滑肌肌动蛋白(α-SMA)的表达,ELISA法检测细胞培养上清液转化生长因子β1(TGF-β1)的含量。结果在一定时间范围内,高糖能刺激细胞Rho A分子活化;与正常对照组比较,葡萄糖60 mmol·L-1培养HK-2细胞72 h后上皮钙黏素表达明显减少(P<0.01),α-SMA表达明显增多(P<0.01),TGF-β1分泌增多(P<0.01),而葡萄糖5.5 mmol·L-1+甘露醇54.5 mmol·L-1高张组无明显差异。法舒地尔20μmol·L-1同步干预后,与高糖组比较,细胞上皮钙黏素表达明显增多〔(0.03±0.01)vs(0.08±0.02),P<0.05〕,α-SMA表达下降(P<0.05),48 h时TGF-β1分泌明显减少〔(128±11)vs(49±5)μg.g-1(P<0.05)〕。结论法舒地尔能抑制高糖培养的肾小管上皮细胞转分化,可能部分通过减少TGF-β1的分泌发挥作用。OBJECTIVE To investigate the effect of fasudil on epithelial-myofibroblast transdifferentiation（EMT） of renal tubular epithelial（HK-2） cells incubated with high concentration of glucose and to explore the mechanism.METHODS HK-2 cells were cultivated with glucose 60 mmol·L-1 and glucose 60 mmol·L-1＋fasudil 20 μmol·L-1.The activity of Rho A was detected with Rho A pull-down assay kit.E-cadherin was detected with immunocytochemical technique and laser confocal scanning microscope.α-Smooth muscle actin（α-SMA）,which reflects the phenotypic characteristics of myofibroblast cells,was detected with Western blotting.Transforming growth factor-β1（TGF-β1） in supernatant was detected with ELISA.RESULTS The activity of Rho A could be activated by high glucose within a certain time.Compared with normal control group,high glucose decreased E-cadherin synthesis but increased α-SMA synthesis（P0.01） and TGF-β1 secretion（P0.01）.There was no differernce between normal control group and glucose 5.5 mmol·L-1＋mannitol 54.5 mmol·L-1 group.Compared with high glucose group,E-cadherin expression was significantly increased（0.03±0.01） vs（0.08±0.02）（P0.05）,α-SMA expression significantly decreased,and TGF-β1 secretion 48 h expression significantly decreased（128±11） vs（49±5）μg·g-1（P0.05） in high glucose＋fasudil group.CONCLUSION Fasudil can inhibit high glucose-induced EMT of renal tubular epithelial cells possibly by reduction of the secretion of TGF-β1.

关 键 词：法舒地尔 糖尿病性肾病 肾小管上皮细胞 

分 类 号：R966[医药卫生—药理学]