肝细胞生长因子抑制高糖诱导肾间质成纤维细胞胶原合成和转化生长因子-β_1的表达  

Hepatocyte growth factor inhibit high glucose induced expression of TGF-β_1 and collagen Ⅲ biosynthesis in interstitial fibroblast

作　　者：牟姗[1] 张庆怡[1] 倪兆慧[1] 赵涵芳[2] 

机构地区：[1]上海第二医科大学附属仁济医院肾内科,上海200001 [2]上海第二医科大学分子生物学研究室

出　　处：《肾脏病与透析肾移植杂志》2004年第1期13-18,共6页Chinese Journal of Nephrology,Dialysis & Transplantation

摘　　要：目的:探讨重组人肝细胞生长因子(rhHGF)对高糖诱导肾间质成纤维细胞损伤的影响。 方法:体外培养肾间质成纤维细胞,在高糖环境下加入rhHGF,以酶联免疫吸附法检测培养上清中型胶原浓度,RT-PCR检测细胞HGF和TGF-b1基因表达,Western blot方法检测TGF-b1蛋白合成。 结果:高糖预处理的细胞型胶原合成增加(同正常对照组相比,P<0.01)。高糖作用早期HGF表达升高,随着作用时间延长,其分泌量下降,而TGF-b1出现持续高表达。此外,HGF可以抑制TGF-b1基因和蛋白表达,结果显示,加入HGF后,TGF-b1基因表达显著下降,表达量下降了50%,并且HGF以剂量依赖形式抑制TGF-b1表达,相关分析证实r = —0.8726,P<0.01。同时HGF对型胶原合成产生抑制作用。 结论:高糖环境促进型胶原合成和TGF-b1持续高表达。rhHGF可以部分抑制型胶原和TGF-b1表达。Objective: Diabetic nephropathy is characterized by unretarded deteriation of glomerular filtration rate clinically and progressive renal fibrosis pathologically. In this study, we investigated the effects of high glucose on interstitial fibroblasts in vitro, probed into the mechanism in which HGF affects on the process of high-glucose induced fibrosis. Methodology: In this study, a cell culture system of human kidney fibroblast (HKF) was used to test high-glucose抯 effects on fibrotic process. The effects of rh-HGF on high-glucose treated HKF was investigated by adding rh-HGF to the medium at different concentration and compared to the control after 48-hours co-culture. Levels of HGF and TGF-b1 were examined at multiple time points with RT-PCR assay. Secreted type  collagen was detected with ELISA. TGF-b1 protein synthesis was detected with Western blot. Results: Production of endogenous HGF mRNA and protein was detected at the sixth hour after HKF incubated in high-glucose culture (vs normal glucose control, P<0.05). Expression of TGF-b1 mRNA and protein was detectable at the 24th hour and mounted to the peak at the 96th hour after HKF incubated in high-glucose culture(vs normal glucose control, P<0.05). Collagen biosynthesis III production was enhanced in high-glucose culture as compared to the normal glucose control. Co-incubation of exogenous rh-HGF with HKF in high-glucose culture for 48 hours decreased TGF-b1 expression in HKF significantly as compared to the control group (P<0.05), and the inhibitive effect of rh-HGF on TGF-b1 protein production was found in a dose-dependant manner. Exogenous rh-HGF also reduced the production of collagen III in HKF by 15% after 48-hours co-incubation (P<0.05). Conclusion: High-glucose can induce TGF-b1 and collagen III biosynthesis in human kidney interstitial fibroblast in vitro, also induce endogenous production of HGF. Exogenous rh-HGF can inhibit high-glucose induced expression of TGF-b1 and collagen Ⅲ biosynthesis in human interstitial fibroblast in v

关 键 词：肝细胞生长因子 肾间质纤维化 成纤维细胞 胶原 转化生长因子-β1 糖尿病肾病 酶联免疫吸附法 

分 类 号：R587.2[医药卫生—内分泌] R692[医药卫生—内科学]