蛋白质乙酰化对HBV复制的影响

Effect of protein acetylation on hepatitis B virus replication

目的探讨宿主细胞蛋白质乙酰化修饰对HBV复制的影响。方法采用去乙酰化酶抑制剂制滴菌素A(TSA)和尼克酰胺(NAM)刺激HBV复制细胞模型Hep G2.2.15和Hep AD38,并检测细胞内的HBV复制指标。采用Western Blot检测广谱乙酰化蛋白和乙酰化组蛋白H3的表达水平。结果 TSA和NAM刺激细胞会引起细胞内蛋白质的乙酰化水平升高,且乙酰化修饰水平增高呈时间和浓度依赖的变化趋势。两种细胞受TSA和NAM刺激的影响,培养上清中的HBs Ag水平降低,而HBV DNA水平升高,且呈时间和浓度依赖关系,而两种细胞培养上清中的HBe Ag和细胞内核心颗粒DNA的表达水平变化趋势不明显。结论宿主蛋白质的乙酰化可能参与和影响了细胞内HBV的复制过程,进一步深入分析和鉴定影响HBV胞内复制的宿主乙酰化蛋白质将有助于加深对HBV复制调控的认识,为开发特异性抗病毒策略提供新思路。

Objective To investigate the effect of protein acetylation in host cells on the replication of hepatitis B virus（ HBV） in hepatocytes,since HBV infection greatly threatens human health and the acetylation of encoding proteins in infected cells plays an important role in HBV replication and infection. Methods The deacetylase inhibitors trichostatin A（ TSA） and nicotinamide（ NAM） were used to stimulate HBV replication in Hep G2. 2. 15 and Hep AD38 cells,and the HBV replication markers were measured. The pan- acetylysin protein and Ac- H3 were examined by Western Blot. Results The stimulation of cells with TSA and NAM increased the overall acetylation level of proteins in cells,and the acetylation level increased in a time- and dose- dependent manner. In the Hep G2. 2. 15 and Hep AD38 cells,stimulation with TSA and NAM reduced HBs Ag level in the supernatant of cell culture and increased HBV DNA level in a time- and dose- dependent manner,while HBe Ag in the supernatant of cell culture and DNA in cells did not change significantly. Conclusion Acetylation of host proteins may be involved in and affect HBV replication in cells,and further analysis and determination of host proteins whose acetylation affects HBV replication in cells help to learn more about the regulation of HBV replication and provide new thoughts for the development of specific antiviral strategies.