表达HBVX基因的小鼠模型的建立及对肝细胞凋亡因子的影响  

Establishment of a mouse model harboring the hepatitis B virus X gene and analysis of expression of apoptosis factors in liver cells of this mouse line

作　　者：郭晨[1] 李丹[1] 林纳[1] 黄月红[1] 陈治新[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化内科,福建省福州市350001

出　　处：《世界华人消化杂志》2011年第12期1225-1230,共6页World Chinese Journal of Digestology

摘　　要：目的:探讨乙型肝炎病毒X(HBVX)基因及其产物对肝细胞凋亡相关基因表达的影响.方法:将实验用SPF级成年♂KM小鼠分为实验组、空质粒对照组、生理盐水对照组.实验组以构建好的PCDNA3.1-HBVX质粒,空质粒对照组以PCDNA3.1质粒,生理盐水对照组以生理盐水,通过尾静脉高压注入动物体内,在第48小时处死小鼠后,取肝组织,以RT-PCR,凝胶回收测序及Westernblot检测HBVX表达,以RT-PCR半定量检测小鼠肝组织内bax、c-myc及bcl-2的表达.结果:实验组RT-PCR显示465bp处有清楚条带,肝组织内有HBVXmRNA的存在,两对照组无HBVXmRNA存在,Western blot检测实验组有HBVX蛋白的表达;两对照组则无HBVX蛋白表达.通过对小鼠肝组织凋亡相关因子的半定量RT-PCR,相对于空质粒对照组和生理盐水对照组,转染HBVX基因的小鼠肝组织bax、c-myc及bcl-2的mRNA相对表达量明显增高,差异有显著性意义(bax:1.3127±0.0900vs1.0023±0.1670,0.9094±0.1081;c-myc:1.6294±0.0672vs1.2869±0.0880,0.9757±0.0397;bcl-2:1.5567±0.1257vs0.6856±0.1554,0.5488±0.1278,均P<0.05).结论:成功建立了表达HBVX的小鼠动物模型,并证明转染HBVX基因后小鼠肝脏组织内bax、bcl-2及c-myc的mRNA表达同时上调.AIM： To investigate the impact of the hepatitis B virus X gene （HBVX） on the expression of apoptosis factors in liver cells.METHODS： The eukaryotic expression vector PCDNA3.1-HBVX was injected into the caudal vein of mice to establish a mouse model expressing the HBVX gene. Mice injected with empty PCDNA3.1 vector or normal saline were used as controls. RT-PCR and Western blot were performed to detect the expression of HBVX in liver tissue 48 h after injection. Semi-quantitative RT-PCR was used to evaluate the expression of bax, bcl-2, and c-myc in liver cells of different groupsof mice.RESULTS： HBX mRNA and protein were detected in liver tissue in mice injected with the PCDNA3.1-HBVX vector, but not in controls. Compared to controls, the expression of bax, c-myc and bcl-2 was up-regulated in liver tissue in mice injected with the PCDNA3.1-HBVX vector （bax： 1.3127 ± 0.0900 vs 1.0023 ± 0.1670, 0.9094 ± 0.1081; c-myc： 1.6294 ± 0.0672 vs 1.2869 ± 0.0880, 0.9757 ± 0.0397; bcl-2： 1.5567 ± 0.1257 vs 0.6856 ± 0.1554, 0.5488 ± 0.1278, all P 0.05）.CONCLUSION： A mouse model harboring the HBVX gene has been established successfully. Expression of the HBx gene can up-regulate the expression of Bax, c-Myc and Bcl-2 in liver cells.

关 键 词：乙型肝炎病毒X基因 肝细胞 凋亡 小鼠模型 

分 类 号：R512.62[医药卫生—内科学]