肝细胞生长因子对人肾小管上皮细胞转分化抑制作用初步研究

Inhibition of Transdifferentiation of Human Renal Tubular Epithelial Cells by Heptocyte Growth Factor

目的 :晚近研究提示肝细胞生长因子 (hepatocytegrowthfactor,HGF)具有减轻实验性肾间质纤维化的作用 ,但其机制尚不十分清楚 ;本研究目的在于探讨HGF对人肾小管上皮细胞 (HKC)转分化的作用。方法 :将体外培养的HKC细胞分为 :(1)无血清培养对照组 ;(2 )阳性对照组 (MCP - 1+AAⅠ ) ;(3)HGF组 (HGF浓度为 0 .0 1,0 .1,1.0 ,10 .0ng/ml) ;(4 )MCP - 1+AAⅠ +HGF组 (HGF浓度 0 .1,1.0 ,10 .0ng/ml)。应用间接免疫荧光法检测HKC细胞内α -平滑肌肌动蛋白 (α -SMA)、波形蛋白、角蛋白表达的变化。应用流式细胞技术检测α -SMA(+)的HKC细胞百分数。结果 :不同浓度HGF分别作用于HKC细胞 48h后 ,经间接免疫荧光法测定该组HKC细胞角蛋白、波形蛋白及α -SMA抗原表达 ,与无血清对照组均无显著差异 ;流式细胞术测得HGF组α -SMA表达阳性HKC细胞百分数 ,与无血清对照组 (平均为 3.1%)也无显著差异 (P >0 .0 5 )。MCP - 1+AAⅠ培养HKC 48h后α -SMA(+)的HKC细胞平均百分数为 85 .6 %。MCP - 1+AAⅠ +HGF(0 .1,1.0 ,10 .0ng/ml)培养HKC 48h后 ,α -SMA(+)的HKC细胞百分数分别为 2 6 .7%,2 .0 %,13.6 %,比阳性对照组显著下降 (P <0 .0 5 )。结论 :(1)HGF对正常HKC细胞转分化无明显影响 ;(2 )

Objective: To examine the possible role of heptocyte growth factor(HGF) in transdifferentiation of human renal tubular epithelial cell lines(HKC).Methods: The HKC cells were divided into(1) negitive control(serum-free);(2)positive control(MCP-1+AAⅠ);(3)HGF group (at doses 0.01,0.1,1.0,10.0 ng/ml);(4)HGF inhibition group(AAⅠ+MCP-1+HGF(at doses 0.1,1.0,10.0 ng/ml).Then the expression of cytocratin,vimentin and α-smooth muscle actin(α-SMA)of HKC cells were assessed by indirect enzyme immuno-histochemistry(EIHC)and flow cytometry.Results: No difference in persentages of α-SMA(+) cells detected by flow cytometry were found between serum-free control and HGF treated ones,treated HKC cells.The expression of vimentin and α-SMA assessed by indirect EIHC in positive controls was markedly increased than negative controls,while the expression of cytocratin was decreased markedly.The expressions of vimentin and α-SMA in MCP-1+AAⅠ+HGF-treated HKC cells were significently decreased than that in positive controls.The persentages of α-SMA(+)HKC cells treated with MCP-1+AAⅠ+HGF(at doses 0.1, 1.0,10.0 ng/ml)were 26.7%,2.0%,13.6%,which was markedly lower than that in positive controls(85.6%,P< 0.005),respectively.Conclusion: (1)There was no transdifferentiating effect when HKC cells cultured with HGF alone.(2)HGF may inhibit the transdifferentiation of HKC cells induced by synergistic effect of MCP-1 and AAⅠ in vitro.