摘要目的 探讨锌α2糖蛋白(AZGP1)在肝星状细胞活化和肝纤维化发生和发展中的作用机制. 方法 采用转化生长因子-β1刺激诱导人肝星状细胞株LX2活化和构建四氯化碳小鼠肝纤维化模型,观察细胞和肝组织中AZGP1的表达情况.应用质粒转染法过表达和抑制表达AZGP1后,分别检测LX2细胞的活化、增殖和凋亡功能及相关因子的改变.多组间比较采用单因素方差分析.结果 免疫荧光染色结果显示在活化的LX2细胞中,AZGP1蛋白减少而α-平滑肌肌动蛋白增多,二者呈负相关.AZGP1基因和蛋白在活化的LX2细胞和四氯化碳肝纤维化小鼠的肝组织内显著低表达.过表达AZGP1后,LX2细胞中Ⅰ型胶原、基质金属蛋白酶-2和α-平滑肌肌动蛋白基因和蛋白明显下调;细胞荧光显示过表达AZGP1的细胞活化和c-平滑肌肌动蛋白减少.此外,过表达AZGP1后,LX2细胞的增殖活性和G1/S-特异性周期蛋白D1蛋白显著降低;细胞周期实验显示过表达AZGP1的细胞停滞在G0/G1期比例显著增多、而S期比例显著减少.AZGP1对LX2细胞凋亡无明显影响. 结论 AZGP1可通过抑制肝星状细胞活化和增殖而逆转肝纤维化;过表达AZGP1有望成为肝纤维化治疗的新靶点.

abstractsObjective To investigate the mechanism of occurrence and development of zinc-alpha-2-glycoprotein (AZGP1) in the activated hepatic stellate cells (HSCs) and liver fibrosis.Methods The activated human hepatic stellate cell line LX2 was induced by the stimulation of transforming growth factor-β1 to construct carbon tetrachloride liver fibrosis mice model.The situation expression of AZGP1 in liver cells and tissues were observed.Plasmid transfection method was used to detect the activation,proliferation,apoptotic functions and changes in related factors of LX2 cells,respectively,after the overexpression and inhibition of AZGP1expression.Univariate analysis of variance was used for multiple group comparison.Results The results of immunofluorescence staining showed that AZGP1 protein was decreased and α-smooth muscle actin was increased in the activated LX2 cells,and the two were negatively correlated.AZGP1 gene and protein were significantly under-expressed in activated LX2 cells and liver tissues of mice with carbon tetrachloride liver fibrosis.Collagen I,matrix metalloproteinase-2,and α-smooth muscle actin genes and proteins were significantly down-regulated in LX2 cells after over-expression of AZGP 1.Cell fluorescence showed that AZGP 1-overexpressing cells were activated and α-smooth muscle actin protein was reduced.In addition,the proliferative activity and G1/S-specific cyclin D1 protein of LX2 cells were significantly reduced after overexpression of AZGP1,while cell cycle experiments showed that the proportion of cells overexpressing AZGP1 was significantly increased in the G0/G1 phase,and the proportion of S phase was significantly reduced.AZGP1 had no significant effect on the apoptosis of LX2 cells.Conclusion AZGP1 can reverse liver fibrosis by inhibiting the activation and proliferation of hepatic stellate cells,and thereby overexpression of AZGP1 is expected to become a new target for liver fibrosis treatment.