小鼠肝脏导入HBx基因诱导MIG的表达

MIG Expression Induced by HBx Gene Delivery into Mouse Liver

目的将HBx基因转入小鼠肝组织并探讨体内HBx对MIG表达的影响。方法携带HBx的质粒和空载体质粒分别与去唾液酸蛋白体内转染系统(in vivo-jetPEI-Gal system)孵育,形成去唾液酸蛋白受体-聚乙烯亚胺-DNA复合物。将复合物经尾静脉高压注射入小鼠体内,2d后处死小鼠摘取肝脏,采用RT-PCR、免疫组织化学、Western blot法分别检测HBx和MIG的表达。结果空白组和阴性对照组小鼠肝组织无HBx表达,实验组RT-PCR测mRNA表达结果为(1.416±0.025),免疫组化法测定HBx蛋白表达阳性面积率为(35.741±2.245)%,Western blot法检测蛋白表达为(1.357±0.031)(P<0.01)。MIG mRNA在空白组、阴性对照组、实验组中的表达分别为(0.106±0.036)、(0.112±0.027)、(0.238±0.051);免疫组化显示MIG蛋白阳性表达面积分别是(10.157±0.841)%、(11.234±0.627)%、(24.568±3.246)%;Western blot检测结果显示MIG蛋白表达分别为(0.149±0.053)、(0.156±0.042)、(0.349±0.087)。MIG mRNA和蛋白在实验组表达均明显高于空白组和阴性对照组(均P<0.05)。结论去唾液酸蛋白与携带HBx质粒形成去唾液酸-聚乙烯亚胺-DNA复合物,通过尾静脉高压注射法可高效地将HBx基因转入小鼠肝脏组织。而且,转入的HBx基因能有效诱导MIG mRNA和蛋白表达。

Objective To deliver HBx gene into mouse liver and investigate MIG expression induced by HBx in vivo.Methods Plasmids containing HBx and control plasmid were incubated with the in vivo-jetPEI-Gal system to produce the asialoglycoprotein receptor-polyethyleneimine-DNA compounds.The in vivo-jetPEI-Gal-DNA compounds were transferred by hydrodynamic injection via the tail vein.The animals were sacrificed and the livers were harvested 2 days following the injection.The expression of HBx and MIG was detected by using RT-PCR,immunohistochemical staining,and Western blot,respectively.Results There was no detectable expression of HBx in mock and negative control groups,and the mRNA and protein expression of HBx in experimental group was（1.416±0.025）,（35.741±2.245） and（1.357±0.031）,respectively（P〈0.01）.Correspondingly,MIG mRNA expression in the mock,negative control and experimental groups was（0.106±0.036）,（0.112±0.027）,（0.238±0.051）,respectively.Moreover,the MIG protein expression in those groups was（10.157±0.841）,（11.234±0.627）,（24.568±3.246） when detected by immunohistochemistry,and was（0.149±0.053）,（0.156±0.042）,（0.349±0.087）,respectively when examined by Western blot.There were statistically significant difference in the MIG expression among three groups（P〈0.05）.Conclusion Hydrodynamic injection of the in vivo-jetPEI-Gal-DNA system containing in vivo-jetPEI-Gal and pCMV-HBx plasmid induces high efficient HBx delivery into mouse liver.Consequently,the exogenous HBx mediates MIG mRNA and protein expression.