K83A变异MxA蛋白抑制HBV复制体外研究

The antiviral activity of K83A mutant MxA protein against the replication of hepatitis B virus in vitro

目的研究K83A变异MxA蛋白抑制HBV复制活性。方法将pcDNA3.1-MxA(wild-type)重组质粒(WT组)、pcDNA3.1-MxA-K83A重组质粒(K83A组)和pcDNA3.1空质粒(对照组)分别与pU19-1.24HBV重组质粒按2∶1比例瞬时共转染HepG2细胞,转染3d后western blot检测MxA蛋白表达,Abbott检测各组细胞上清HBsAg和HBeAg表达,定量PCR检测上清和细胞内HBV DNA水平,统计学分析结果。结果MxA、K83A组有较好的MxA蛋白表达;与对照组相比,K83A组和MxA组上清HBeAg分别下降73%和71%(P<0.05),上清HBV DNA水平分别下降2.22lg和2.11lg(P<0.01),细胞内HBV DNA水平下降1.89lg和1.78lg(P<0.01)。结论K83A变异不影响MxA蛋白抑制HBV复制活性。

Objective To investigate the antiviral activity of K83A mutant MxA protein against the replication of hepatitis B virus. Methods The recombinant vectors of pcDNA3, 1 -MxA （wildtype）, pcDNA3. 1 -MxA（K83A） were cotransfeeted with PU19 - 1, 24HBV repectively into HepG2 cells. After 3 dasys,the expression of MxA protein was detected by western blot. The superuatant HBsAg and HBeAg were measured by abbott analysis. HBV DNA was detected by real - time PCR. Results The MxA protein was expressed robustly in HepG2 cells of WT and K83A groups detected by western blot. Compared with the control group,the superuatant HBeAg in WT and K83A groups was decreased by 73% and 71% ,the extracellular HBV DNA was decreased by 2. 22 lg and 2. 11 lg respectively and the intracellnlar HBV DNA was decreased by 1.89 lg and 1.78 lg respectively. Conclusion K83A mutation shows no influence on the interferon - inducible MxA protein to inhibit HBV replication.