强的松增强 HBV 感染小鼠体内病毒复制表达

目的:检测糖皮质激素对HBV感染小鼠体内HBV复制的作用。方法:采用高压注射体内转染法将HBV复制型重组质粒p AAV/HBV1.2导入雄性C57BL/6小鼠,转染后24 h按体重、年龄、血清HBe Ag水平对小鼠进行配对,分为实验组和对照组,分别连续给予强的松或生理盐水灌胃。于第3天和第10天给药后4~6 h处死小鼠。采用Southern杂交检测小鼠肝脏HBV-DNA复制中间体;化学发光法检测血清中HBe Ag和HBs Ag。结果:对HBV感染小鼠连续给予强的松或生理盐水3 d后,所有小鼠肝脏中均可检测到较高水平的HBV-DNA复制中间体,给予强的松的实验组稍高于给予生理盐水的对照组,二者比值为1.127∶1;连续处理10 d后,实验组小鼠肝脏中HBV-DNA复制中间体的水平明显高于对照组,比值为2.534∶1。处理3 d后对照组和实验组小鼠血清HBs Ag平均值分别为:1 618.28μg/μL、1 666.23μg/μL,实验组与对照组无明显差别;小鼠血清对照组和实验组HBe Ag平均值分别为14.68μg/μL、14.95μg/μL,实验组对照组无明显差别;处理10 d后对照组和实验组小鼠血清HBs Ag平均值分别为:8 059.40μg/μL、11 885.87μg/μL,实验组高于对照组。小鼠血清对照组和实验组HBe Ag平均值分别为183.15μg/μL、286.14μg/μL,实验组高于对照组。结论:强的松可增强HBV感染小鼠体内乙肝病毒的复制和表达,并且随着作用时间的延长,作用也越明显。本研究为糖皮质激素及引起HBV再激活的相关研究奠定基础,同时也再次提醒临床医生,应充分重视糖皮质激素的使用中HBV再激活的问题。

Prednisone enhancing viral replication and expression in HBV transfection mice

Objective:To detect the effect of glucorticoid on HBV replication of mice in vivo.Methods:HBV replication com-petent plasmid pAAV /HBV1 .2 was transferred into male C57L /6 mice by hydrodynamics-based in vivo transfection.24 h after transfec-tion,the mice were matched by body weight,age and serum level of Hepatitis B e antigen (HBeAg)and were divided into experiment group and control group,and then treated orally with prednisone or normal saline at interval of twenty four hours for 3 and 10 days.Four to six hours after the last administration,the mice were killed.HBV-DNA replication intermediates in liver were analyzed by Southern Blot Hybridization.The serum HBsAg and HBeAg were detected by chemiluminescent detection system.Results:HBV-DNA replication intermediates were detected in the livers of mice after treating orally with prednisone or normal saline for 3 days,the level of HBV-DNA replication intermediates of the experimental group was 0.127 times higher than that of the control group.However,after treatment for 10 days,compared with the control group of mice,the level of HBV-DNA replication intermediates of the experimental mice increased greatly.Quantitative analysis showed that prednisone enhanced the level of HBV-DNA replication intermediates appropriately 1 .534 fold.Three days after the above treatment,the serum HBsAg of the control mice and the experimental mice were 1 618.28 μg /μL and 1 666.23 μg /μL on average respectively,which showed no significant difference.Similarly,there was no significant difference in the se-rum HBeAg,which showed as follows:14.68 μg /μL,14.95 μg /μL on average respectively.In contrast,10 days after the same treat-ment,the serum HBsAg of the control and the experimental of mice were on average 8 059.40 μg /μL,1 1 885.87 μg /μL,respectively, and the serum HBeAg were 183.15 μg /μL,286.14 μg /μL,respectively.Both of the level of serum HBsAg and HbeAg from experi-mental mice were higher than that of control animals.Conclusion:The replication and expression of hepatitis B virus in the HBV infec-tion mice could be increased by prednisone orally and the increasing effect of prednisone on HBV is time-dependant,which provides fur-ther research about HBV reactivation caused by prednisone and other immunosuppressive drugs,and reminds clinicians,especially non-infected professional doctors to pay full attention to HBV reactivation when glucocorticoid is used in patients with HBV infection.