丙型肝炎病毒F蛋白诱导LX2肝星形细胞活化

Activation of hepatic stellate LX2 cells by F protein of hepatitis C virus

目的探讨丙型肝炎病毒(HCV)F蛋白对肝星形细胞(HSC)功能的影响。方法采用脂质体转染法将pcDNA3.1-f(实验组)或pcDNA3.1空质粒(对照组)转染HSC株LX2细胞,24、48 h后收集细胞,用Western blot法检测α-肌动蛋白(α-SMA)含量,RT-PCR法测定金属蛋白酶组织抑制因子1(TI MP-1)和基质金属蛋白酶2(MMP-2)基因mRNA表达水平。结果转染后24、48 h,实验组LX2细胞α-SMA蛋白表达量以及TI MP-1基因mRNA水平均明显高于对照组,两组细胞MMP-2基因mRNA水平接近。结论 HCV F蛋白能够活化HSC,上调其TI MP-1基因转录水平,但不影响MMP-2基因转录水平,使MMP-2蛋白的有效酶活性下降,造成肝组织细胞外基质(ECM)降解速率减慢,进而发生纤维化。

Objective To investigate the effect of hepatitis C virus（HCV） F protein on function of hepatic stellate cells.Methods 24 h and 48 h after transfection pcDNA3.1-f（test group）or pcDNA3.1（control group） plasmid into hepatic stellate LX2 cells by liposome,the protein of smooth muscle α-actin（α-SMA） was detected by Western blot,and the mRNA level of tissue inhibitor factor 1 of metalloproteinases（TIMP-1） and matrix metalloproteinase 2（MMP-2） gene were examined by RT-PCR.Results 24 h and 48 h after LX2 cells transfected pcDNA3.1-f（test group） or pcDNA3.1（control group）,the content of α-SMA protein and the mRNA level of TIMP-1 gene of cells in test group was high than that in control group,and the mRNA level of MMP-2 gene of cells showed no difference between test and control groups.Conclusion HCV F protein is able to activate hepatic stellate LX2 cells,enhance the mRNA level of TIMP-1 gene,but not affect MMP-2 gene transcription,which make the effective enzyme activity of MMP-2 protein be decreased,the degradation rate of ECM in hepatic tissue slowed down,and result in hepatic fibrosis.