阿维霉素干预MTIF2调控核糖体翻译进程抑制乙型肝炎病毒复制的机制研究

A study on the mechanism of Avilamycin intervention MTIF2 regulating ribosomal translation process to inhibit hepatitis B virus replication

目的探讨核糖体翻译因子抑制剂阿维霉素对乙型肝炎病毒复制的抑制作用及分子机制。方法使用不同浓度的阿维霉素处理肝癌细胞Hep3B,应用CCK-8检测细胞活性,流式细胞法检测细胞凋亡,qPCR方法检测病毒HBV-DNA、pgRNA、宿主MTIF2和RPL10基因表达量,ELISA免疫测法检测HBsAg和HBeAg,化学发光检测法对AFP进行定量检测,生物化学方法检测细胞外泌谷草转氨酶(AST)、谷丙转氨酶(ALT)和碱性磷酸酶(ALP)蛋白。结果阿维霉素对Hep3B细胞增殖没有抑制作用,对细胞的凋亡也没有影响。但能促进细胞AST分泌,降低AFP外泌水平,对ALP分泌影响较小。在Hep3B细胞中,阿维霉素能通过干预MTIF2促使pgRNA表达量积累,反馈上调宿主RPL10和MTIF2基因mRNA表达。并能有效降低HBsAg、HBeAg和HBV-DNA水平。结论阿维霉素能够抑制MTIF2翻译起始,通过影响翻译起始调控病毒组装蛋白的翻译进程,进而抑制乙肝病毒复制。

Objective To investigate the inhibitory effect and molecular mechanism of ribosomal translation factor inhibitor Avilamycin on hepatitis B virus replication.Methods Liver cancer Hep3B cells were treated with different concentrations of Avilamycin.Cell activity was detected by CCK-8;the apoptosis was detected by flow cytometry,and HBV-DNA、pgRNA、MTIF2、RPL10 gene expression level was detected by qPCR method.The HBsAg and HBeAg was detected by ELISA.The AFP was detected by chemiluminescence.Aspartate aminotransferase(AST),alanine aminotransferase(ALT),and alkaline phosphatase(ALP)proteins was detected by Biochemistry method.Results Avilamycin had no inhibitory effect on Hep3B cell proliferation and apoptosis.However,it could promote cellular AST secretion,reduce AFP levels,and have less effect on ALP secretion.In Hep3B cells,Avilamycin promotes accumulation of pgRNA expression by intervening with MTIF2 and feedback upregulates mRNA expression of host RPL10 and MTIF2 genes.It can effectively reduce the HBsAg,HBeAg,and HBV-DNA levels.Conclusion Avilamycin can inhibit MTIF2 translation initiation,regulate the translation process of viral assembly protein by affecting translation initiation,and then inhibit hepatitis B virus replication.