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目的观察促肝细胞生长素(hepatocyte growth-promoting factor,pHGF)对巨噬细胞趋化因子(monocyte chemo-tatic protein-1,MCP-1)协同马兜铃酸Ⅰ(aristolochic acidⅠ,AAⅠ)诱导的人肾小管上皮细胞(HKC)凋亡及上皮细胞-间质细胞转分化(epithelial-mesenchymal transition,EMT)的影响。方法体外培养的HKC随机分为:空白对照组、转分化模型组及不同浓度pHGF(0.15、1.5、15、150、1500ng/ml)处理组。转分化模型组采用MCP-1(0.1μg/ml)协同AAⅠ(10μg/ml)诱导HKC转分化模型;pHGF处理组采用不同浓度pHGF对转分化模型HKC进行处理;空白对照组常规培养。采用WST-8法和流式细胞术观察各组细胞增殖和凋亡情况;RT-PCR检测各组细胞α-SMA mRNA表达;免疫组化检测各组细胞α-SMA、TGF-β1、FN蛋白的表达。结果与空白对照组相比,转分化模型组、不同浓度pHGF处理组HKC细胞增殖抑制率,凋亡细胞所占比例,α-SMA mRNA表达均明显升高(P<0.01),提示转分化模型制备成功。与转分化模型组细胞相比,pHGF(150ng/ml)处理组HKC增殖抑制率明显降低(P<0.01),各浓度pHGF处理组HKC凋亡细胞所占比例均明显降低(P<0.01),HKC细胞α-SMA mRNA表达下调(150ng/ml pHGF处理组尤明显);α-SMA、TGF-β1、FN蛋白表达下调。结论pHGF(150ng/ml)可部分逆转MCP-1协同AAⅠ诱导的HKC增殖抑制、凋亡和EMT。
Objective To investigate the effects of hepatocyte growth-promoting factor (pHGF) on macrophage chemo-tatic protein-1 (MCP-1) and aristolochic acid Ⅰ (AAⅠ) Apoptosis of human renal tubular epithelial cells (HKC) and epithelial-mesenchymal transition (EMT). Methods HKCs cultured in vitro were randomly divided into control group, transdifferentiation model group and different concentrations of pHGF (0.15,1.5,15,150,1500 ng / ml) treatment group. HKCs transdifferentiation model was induced by MCP-1 (0.1μg / ml) in combination with AAⅠ (10μg / ml) in transdifferentiation model group. HKCs treated with different concentrations of pHGF in pHGF treatment group were cultured in blank control group. The WST-8 method and flow cytometry were used to observe the cell proliferation and apoptosis in each group. The expression of α-SMA mRNA in each group was detected by RT-PCR. The expressions of α-SMA, TGF-β1 and FN expression. Results Compared with the blank control group, the proliferation inhibition rate, the percentage of apoptotic cells and the expression of α-SMA mRNA in HKC cells treated with different concentrations of pHGF were significantly increased (P <0.01), indicating that the transdifferentiation model Preparation is successful. Compared with the transdifferentiation model group, the proliferation inhibition rate of HKC in the group treated with pHGF (150ng / ml) was significantly decreased (P <0.01), and the percentage of apoptotic cells in HKC treated group was significantly decreased (P <0.01) The expression of α-SMA mRNA in HKC cells was down-regulated (especially in 150ng / ml pHGF treatment group), and the expressions of α-SMA, TGF-β1 and FN were down-regulated. Conclusions pHGF (150ng / ml) can partly reverse the proliferation inhibition, apoptosis and EMT induced by MCP-1 in combination with AA Ⅰ.