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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u001f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u001f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u001f型胶原合成产生抑制作用。 结论:高糖环境促进u001f型胶原合成和TGF-b1持续高表达。rhHGF可以部分抑制u001f型胶原和TGF-b1表达。
Objective: To investigate the effect of recombinant human hepatocyte growth factor (rhHGF) on the injury of renal interstitial fibroblasts induced by high glucose. METHODS: Human renal interstitial fibroblasts were cultured in vitro. RhHGF was added in high glucose. The concentration of collagen in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The expression of HGF and TGF-β1 was detected by RT-PCR. blot method to detect TGF-b1 protein synthesis. Results: The hyperglycemic cells pretreated with u001f collagen increased (P <0.01 compared with the normal control group). In the early stage of high glucose, the expression of HGF was increased. With the prolongation of action time, the secretion of HGF was decreased, while the expression of TGF-b1 was continuously high. In addition, HGF can inhibit TGF-β1 gene and protein expression, the results showed that the addition of HGF, TGF-b1 gene expression decreased significantly, the expression decreased by 50%, and HGF dose-dependent inhibition of TGF-b1 expression, the correlation analysis confirmed r = -0.8726, P <0.01. At the same time HGF on u001f collagen synthesis inhibition. CONCLUSION: High glucose environment can promote collagen synthesis and TGF-b1 expression continuously. rhHGF can partially inhibit the expression of u001f collagen and TGF-b1.