辛伐他汀抑制重组人偏肺病毒复制的自噬作用机制研究

Research of simvastatin-induced autophagy in inhibiting recombinant human metapneumovirus replication

目的:探讨人偏肺病毒(human metapneumovirus,hMPV)感染后辛伐他汀是否可以抑制病毒复制。方法:体外试验,hMPV感染16HBE细胞(人支气管上皮样细胞)后用辛伐他汀试剂处理,qPCR和Western blot检测病毒滴度和自噬的强弱以及相关通路表达水平;体内试验,利用辛伐他汀灌胃处理被hMPV感染的BALB/c小鼠,肺组织切片观察病理情况,提取肺组织蛋白和RNA检测病毒载量的变化、自噬发生情况以及相关通路的表达。结果:体外试验,辛伐他汀干预组病毒滴度降低,自噬水平高于病毒组;辛伐他汀干预组AKT/mTOR通路被抑制,使用通路特异性抑制剂雷帕霉素处理后进一步验证了通路的准确性。体内试验,辛伐他汀干预组病毒滴度低于病毒组,但自噬表达水平两组无明显差异,辛伐他汀干预组AKT/mTOR通路下调;HE染色病毒组病理改变明显,辛伐他汀干预后病理情况改善。结论:辛伐他汀在人偏肺病毒感染后可以抑制其复制,该过程与AKT/mTOR通路诱导的自噬反应相关。

Objective To study whether simvastatin could inhibit viral replication during human metapneumovirus(hMPV)infection.Methods Human bronchial epithelial cells(16HBE)were infected with hMPV and then treated with or without simvastatin.Real-time quantitative PCR(qPCR)and Western blot were used to detect virus titers and the activation of autophagy and related pathways.BALB/c mice were infected with hMPV and then treated with simvastatin through intragastric administration.Pathological changes in lung tissues were observed.Changes in viral loads and the activation of autophagy and related pathways in proteins and RNA extracted from lung tissues were detected.Results The in vitro experiment showed that the hMPV+simvastatin group had decreased virus titer and enhanced autophagy than the hMPV group.The AKT/mTOR pathway in the hMPV+simvastatin group was inhibited,which was verified by a further experiment using rapamycin,a specific inhibitor of AKT/mTOR pathway.The in vivo experiment showed that the virus titer in the hMPV+simvastatin group was lower than that in the hMPV group,but there was no significant difference in the activation of autophagy.The AKT/mTOR pathway was down-regulated in the hMPV+simvastatin group.HE staining revealed that obvious pathological changes were observed in the hMPV group,but the condition was improved after simvastatin intervention.Conclusions Simvastatin can inhibit the replication of hMPV,which is associated with the activation of autophagy induced by AKT/mTOR pathway.