Luo S Z, Wu J C, Lin R K, Liu T, Luo X, Xu M Y, Guo C Y
Department of Gastroenterology and Hepatology, Shanghai Tenth People's Hospital of Nanjing Medical University, Shanghai 200080, China; Department of Gastroenterology and Hepatology, Shanghai General Hospital, Shanghai Jiaotong University of Medicine, Shanghai 200080, China.
Department of Gastroenterology and Hepatology, Shanghai General Hospital, Shanghai Jiaotong University of Medicine, Shanghai 200080, China.
Zhonghua Gan Zang Bing Za Zhi. 2020 Feb 20;28(2):135-140. doi: 10.3760/cma.j.issn.1007-3418.2020.02.007.
To investigate the mechanism of occurrence and development of zinc-alpha-2-glycoprotein (AZGP1) in the activated hepatic stellate cells (HSCs) and liver fibrosis. 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. 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 AZGP1. Cell fluorescence showed that AZGP1-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. 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.
探讨锌-α-2-糖蛋白（AZGP1）在活化肝星状细胞（HSCs）及肝纤维化发生发展中的作用机制。采用转化生长因子-β1刺激诱导活化人肝星状细胞系LX2，构建四氯化碳肝纤维化小鼠模型。观察肝细胞及组织中AZGP1的表达情况。分别采用质粒转染法检测AZGP1表达上调和抑制后LX2细胞的活化、增殖、凋亡功能及相关因子变化。采用单因素方差分析进行多组比较。免疫荧光染色结果显示，活化的LX2细胞中AZGP1蛋白减少，α-平滑肌肌动蛋白增加，二者呈负相关。AZGP1基因和蛋白在四氯化碳肝纤维化小鼠的活化LX2细胞及肝组织中显著低表达。AZGP1过表达后，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及蛋白显著下调。细胞荧光显示，过表达AZGP1的细胞活化，α-平滑肌肌动蛋白蛋白减少。此外，AZGP1过表达后，LX2细胞的增殖活性及G1/S特异性细胞周期蛋白D1蛋白显著降低，但细胞周期实验显示，过表达AZGP1的细胞在G0/G1期比例显著增加，S期比例显著降低。AZGP1对LX2细胞凋亡无显著影响。AZGP1可通过抑制肝星状细胞的活化和增殖来逆转肝纤维化，因此AZGP1的过表达有望成为肝纤维化治疗的新靶点。
