蛋白酶体抑制剂PS-341抑制人肠道病毒D68复制研究  

Proteasome inhibitor bortezomib inhibits replication of Enterovirus D68

作　　者：张可可 夏冬[2] 刘思华 周振威 韩俊 王涛 Zhang Keke;Xia Dong;Liu Sihua;Zhou Zhenwei;Han Jun;Wang Tao(School of Life Sciences, Tianjin University, Tianjin 300110;National Institute for Viral Disease Control and Prevention , Chinese Center for Disease Control and Prevention, State Key Laboratory of Infectious Disease Prevention and Cotrol, Collaborative Innovation Center for Diagnosis and Treatment of Infections Disease, Beijing 102206, China)

机构地区：[1]天津大学生命科学学院,300110 [2]中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室传染病诊断和治疗协同创新中心,北京1002206

出　　处：《中华实验和临床病毒学杂志》2019年第3期236-243,共8页Chinese Journal of Experimental and Clinical Virology

基　　金：国家重点研发项目(2017YFA0205102).

摘　　要：目的探究蛋白酶体抑制剂PS-341对肠道病毒属病毒复制的抑制作用。方法利用MTT实验分析药物毒性。利用实时荧光定量PCR技术,从核酸水平检测PS-341处理对宿主细胞内肠道病毒 D68 型(enterovirus D68 ,EV-D68)、柯萨奇 B3 型(coxsackieviros B3 , CV-B3 )复制的影响。通过Western blot检测细胞内病毒蛋白表达水平。结果 PS-341处理EV-D68或CV-B3感染后的细胞,细胞内病毒RNA含量分别被抑制50%?70%和60%~90%。EV-D68感染细胞后,加入PS-341, RD细胞内病毒滴度下调90.23%,上清中病毒滴度下调83.4%,HeLa细胞内病毒滴度下调93.08%,上清中病毒滴度下调90%。但PS-341对病毒吸附、进入及释放无影响。当PS-341与凋亡抑制剂Ac-YVADCHO共处理EV-D68感染的细胞,与PS-341处理组相比,PS-341对病毒RNA复制抑制率为10%~30%,同时病毒蛋白表达水平提高,PS-341对病毒复制抑制作用减弱。结论 PS-341能够抑制肠道病毒属病毒在细胞内复制及组装,但对病毒吸附、进入及释放无影响。PS-341可能通过调节蛋白酶体通路,诱导细胞凋亡,抑制EV-D68在宿主体内的基因复制抑制病毒增殖。此外,PS-341对小RNA病毒科肠道病毒属病毒可能具有广谱抗病毒作用。Objective To investigate the inhibitory effect of bortezomib ( PS-341 ) on enterovirus replication. Methods The methyl thiazolyl tetrazolium ( MTT) assay was used to value cell viability in response to PS-341 treatment. The protein and viral gene mRNAs were measured by real-time quantitative PCR ( qRT-PCR). Results Our result show that after enterovirus ( EV)-D68 or coxsackievirus B3 ( CVB3) infected cells were treated with PS-341 , compared with the control group, the inhibition rate of the intracellular viral RNA reached 50%~ 70% or 60%~ 90%. PS-341 was added after RD cells were infectd with EV-D68, the intracellular virus titer was down-regulated by 90. 23% and 83. 40% in the supernatant, the intracellular virus titer was down-regulated by 93% and 90% in the supernatant and in RD cells. PS-341 had no effect on virus adsorption and importing. The cells were treated with PS-341 and apoptosis-inhibiting agent Ac-YVAD-CHO, the viral RNA replication inhibition rate reached 10%-30%, and the expression of viral protein was increased, which indicated that the inhibitory effect of PS-341 on viral replication wasattenuated. Conclusions According to the result of the study, PS-341 could reduce apoptosis by regulating the proteasome pathway, inhibiting the gene replication and assemble, without effect on virus adsorption, entry and release. In addition, PS-341 also inhibited the replication of CV-B3 in cells, which suggest that PS?341 has a broad spectrum anti-EVs effects.

关 键 词：肠道病毒D68 柯萨奇病毒B3 PS-341 细胞凋亡 

分 类 号：R373.2[医药卫生—病原生物学]