| 摘要: |
| 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 chemotatic protein-1,MCP-1) 协同马兜铃酸Ⅰ(aristolochic acid Ⅰ,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、1 500 ng/ml)处理组。转分化模型组采用MCP-1(0.1 μg/ml)协同AAⅠ(10 μg/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(150 ng/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(150 ng/ml pHGF处理组尤明显)；α-SMA、TGF-β1、FN蛋白表达下调。结论pHGF(150 ng/ml)可部分逆转MCP-1协同AAⅠ诱导的HKC增殖抑制、凋亡和EMT。 |
| 关键词: 促肝细胞生长素 肾小管上皮细胞 细胞凋亡 上皮-间质细胞转分化 |
| DOI：10.3724/SP.J.1008.2010.051 |
| 投稿时间：2009-02-24修订日期：2009-12-09 |
| 基金项目:广州市科技计划项目(2002 J1-C0361),广东省医学科研基金(A2002349). |
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| Hepatocyte growth-promoting factor partially reverses monocyte chemotatic protein-1- and aristolochic acidⅠ-induced epithelial-to-mesenchymal transition of human kidney epithelial cells |
| CHEN Chao1,3,JIANG Jian-wei2*,ZHOU Xu-long1,YAN Yu-xia2,CHEN Tao2,ZHANG Xiao-ying2 |
| (1.Department of Pathology,Medical College,Jinan University,Guangzhou 510632,Guangdong,China;2.Department of Biochemistry,Medical College,Jinan University,Guangzhou 510632,Guangdong,China;3.Department of General Surgery,the People’s Hospital of Jiaozuo City,Jiaozuo 454002,Henan,China) |
| Abstract: |
| ObjectiveTo observe the influence of hepatocyte growth-promoting factor (pHGF) on monocyte chemotatic protein-1-(MCP-1) and aristolochic acid Ⅰ(AAⅠ)-induced epithelial-to-mesenchymal transition (EMT) and apoptosis of human kidney epithelial cell line(HKC).MethodsThe HKC cells were randomly divided into blank control group (control groups), epithelial-to-mesenchymal transition model group (model group),and pHGF inhibition group (pHGF groups) with pHGF at different concentrations (0.15,1.5,15,150,and 1 500 ng/ml).The EMT model was established by exposing HKC cells to MCP-1(0.1 μg/ml)and AAⅠ(10 μg/ml).Cells in the pHGF groups were the model cells treated with different concentrations of pHGF.Cells in the control group were cultured routinely.WST-8 method and flow cytometry were used to observe the proliferation and apoptosis of HKC cells,respectively.The mRNA expression of α-smooth muscle actin(α-SMA) was determined by reverse transcriptase-polymerase chain reaction(RT-PCR),and the expression of α-SMA,fibronectin (FN),and transforming growth factor-β1(TGF-β1) in HKC cells were assessed by indirect enzyme immunohistochemistry.ResultsThe cell inhibitory rate,apoptotic rate,and expression of α-SMA mRNA were significantly increased in the model group and pHGF groups compared with those in the control group (P<0.01),indicating the successful establishment of EMT model.Compared with the model group,pHGF at 150 ng/ml,but not at other concentrations,significantly decreased the inhibition rate of HKC cells(P<0.01).The apoptotic rate of HKC cells in all the pHGF groups were significantly lower than that in the model group (P<0.01).pHGF at 150 ng/ml also greatly decreased the expression of α-SMA mRNA,and significantly down-regulated the expression of α-SMA,TGF-β1,and FN protein.ConclusionpHGF at 150 ng/ml can partly reverse MCP-1- and AA Ⅰ-induced HKC cell growth inhibition,apoptosis,and EMT. |
| Key words: hepatocyte growth-promoting factor kidney epithelial cells apoptosis epithelial-to-mesenchymal transition |
