论文部分内容阅读
目的检测HBV preS1蛋白及ASGPR的亚细胞定位及在肝组织中的分布情况,为研究二者的生物学功能提供依据。方法激光共聚焦显微镜检测HBV preS1蛋白及ASGPR在HepG2细胞中的亚细胞定位情况以及二者是否存在共定位;免疫组织化学技术检测HBV preS1蛋白和ASGPR在肝组织中的表达情况。结果激光共聚焦显微镜下观察,在HepG2细胞的胞膜、胞质及胞核中均可见明亮的绿色荧光,表明HBV preS1蛋白弥散分布于肝细胞的胞膜、胞质及胞核中;在胞膜上可见明亮的红色荧光,而胞质中的红色荧光较弱,表明ASGPR主要表达于肝细胞膜上,在肝细胞质中呈弱表达。提示在HepG2细胞的细胞膜上HBV preS1蛋白与ASGPR具有共定位。在乙肝肝硬化患者的肝组织中,HBV preS1蛋白在肝细胞的胞膜、胞质及胞核中均有阳性表达;ASGPR不仅在肝细胞膜上有阳性表达,在胞质中的表达率也较高。此外,ASGPR在肝组织中的分布形式与HBV preS1蛋白无明显差异(P>0.05),并且二者的表达水平呈显著正相关(Pearson相关系数0.776,P<0.000 1)。结论 ASGPR除介导HBV入侵外,可能还参与了病毒的分泌及胞内运输等其他生命活动。
Objective To detect subcellular localization of HBV preS1 protein and ASGPR and its distribution in liver tissue, and to provide basis for the study of their biological functions. Methods The subcellular localization of HBV preS1 protein and ASGPR in HepG2 cells was detected by laser confocal microscopy. The coexpression of HBV preS1 protein and ASGPR in liver tissues was detected by immunohistochemistry. Results Under laser scanning confocal microscope, bright green fluorescence was observed in the membrane, cytoplasm and nucleus of HepG2 cells, indicating that HBV preS1 protein was diffusely distributed in the membrane, cytoplasm and nucleus of hepatocytes. Bright red fluorescence was seen on the membrane, whereas the red fluorescence in the cytoplasm was weak, indicating that ASGPR was mainly expressed on the hepatocyte membrane and weakly expressed in the liver cytoplasm. Suggesting that HBV preS1 protein co-localized with ASGPR on the cell membrane of HepG2 cells. In the liver tissues of patients with hepatitis B cirrhosis, HBV preS1 protein was positively expressed in the membrane, cytoplasm and nucleus of hepatocytes. ASGPR not only expressed in the liver cell membrane but also in the cytoplasm high. In addition, the distribution of ASGPR in liver tissue was not significantly different from HBV preS1 protein (P> 0.05), and the expression level of ASGPR was positively correlated (Pearson correlation coefficient 0.776, P <0.0001). Conclusion In addition to mediating HBV invasion, ASGPR may also be involved in other life activities such as secretion of the virus and intracellular transport.