摘要
目的观察乙型肝炎病毒(HBV)感染对小鼠体内视黄醇结合蛋白4(RBP4)表达的影响。方法将C57BL/6小鼠随机分为正常组,HBV感染组(尾静脉高压注射rAAV8-1.3HBV病毒载体1×10^11 vg/只),HBV感染+拉米夫定喂养组(拉米夫定150 mg/kg),分别于6周、12周和18周麻醉后处死6只小鼠。荧光定量PCR(RT-qPCR)检测血清HBV DNA含量;全自动生化仪以及ELISA检测血清相关生化指标以及RBP4含量;ELISA检测肝脏组织中肿瘤坏死因子α(TNF-α)以及白细胞介素6(IL-6)炎性因子水平;RT-qPCR、Western blot分别检测肝脏组织中RBP4 mRNA以及蛋白的表达。结果HBV感染组小鼠血清中HbsAg均保持较高水平且阳性率稳定在80%以上,小鼠血清HBV DNA含量显著高于对照组(P<0.05),HBV感染小鼠模型构建成功;感染组小鼠肝脏组织中TNF-α以及IL-6水平均随时间的增加而升高,其中第12周、18周时要显著高于正常组(P<0.05);此外,感染组小鼠肝脏组织中RBP4蛋白以及RBP4 mRNA的表达水平均随感染时间的增加而下降,其水平在第6周、12周、18周均要低于正常组以及拉米夫定喂养组(P<0.05)。结论HBV感染小鼠后能够显著降低小鼠肝脏组织中视黄醇结合蛋白4的表达水平。
Objective To observe the effect of hepatitis B virus(HBV)infection on the expression of retinol binding protein 4(RBP4)in mice.Methods C57BL/6 mice were randomly divided into normal group,HBV infected group(vail vein hydrodynamic injectionr AAV8-1.3HBV viral vector1×10 11 vg/per)and HBV infected+lamivudine group(150 mg/kg).Then 6 mice were sacrificed after 6,12 and 18 weeks of injection.The level of HBV DNA in serum was detected by RT-qPCR.Then,the RBP4 and biochemical indicators in mice serum were detected by ELISA and automatic biochemical analyzer;the levels of inflammatory factors(TNF-α,IL-6)in liver tissues were detected by ELISA,and the expression of RBP4 protein and RBP4 mRNA in liver tissue were detected by RT-qPCR.Results The HBV-infected mice were successfully constructed,the serum HbsAg remained in high level and the positive rate was stable with more than 8%at 1-18 weeks after the tail vein hydrodynamic injection,and the HBV DNA in serum of mice was also significantly higher than that in the control group.The levels of TNF-αand IL-6 in the liver tissue from the infected mice increased with time,which was significantly higher than that in the normal group at 12 and 18 weeks after injection(P<0.05).In addition,the expression level of liver RBP4 protein and RBP4 mRNA from the infected group decreased over infection time,which was lower than that in the normal group and the lamivudine group at the 6,12 and 18 weeks after injection(P<0.05).Conclusions Infection of HBV can significantly inhibit the expression level of liver retinol binding protein 4 in mice.
作者
何邵波
伍美兰
李小民
李丽钱
HE Shao-bo;WU Mei-lan;LI Xiao-min;LI Li-qian(School of Medical Laboratory,Shaoyang University,Shaoyang 422000;Department of Clinical Laboratory,the Second Affiliated Hospital of Shaoyang University,Shaoyang 422000;Department of Clinical Laboratory,Xinshao County People's Hospital,Shaoyang 422900,China)
出处
《基础医学与临床》
CSCD
2019年第9期1277-1282,共6页
Basic and Clinical Medicine
基金
湖南省教育厅科学研究一般项目(14C1037)
