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

作　　者：唐莹莹[1] 田德安[1] 夏雨佳[1] 晏维[1] 张全乐[1] 王伟[1] 常莹[1] 刘梅[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院消化内科,武汉430030

出　　处：《华中科技大学学报（医学版）》2012年第4期395-399,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30972759)

摘　　要：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.

关 键 词：HBX MIG 去唾液酸蛋白受体 高压水注射法 

分 类 号：R575.1[医药卫生—消化系统]