牛蒡苷元通过调控宿主基因表达和细胞凋亡抑制寨卡病毒复制的研究  

Arctigenin Inhibits Zika Virus Replication by Modulating Host Gene Expression and Cell Apoptosis

作　　者：陈雨露 冯燕[2] 张哲雯 徐昌平[2] 吴巧凤[1] CHEN Yulu;FENG Yan;ZHANG Zhewen;XU Changping;WU Qiaofeng(School of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053;Zhengjiang Provincial Center for Disease Control and Prevention,Hangzhou 310051)

机构地区：[1]浙江中医药大学药学院,杭州310053 [2]浙江省疾病预防控制中心,杭州310051

出　　处：《中药药理与临床》2023年第4期40-46,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：浙江省自然科学基金(编号:LY18H280007)。

摘　　要：目的:探讨牛蒡苷元抗寨卡病毒(Zika virus, ZIKV)的药效及其作用机制。方法:采用qRT-PCR及CPE法考察在非洲绿猴肾(Africa green monkey kidney cells, Vero)细胞上,牛蒡苷元抗ZIKV的活性及作用方式;转录组测序(RNA-sequencing, RNA-seq)分析正常Vero细胞、ZIKV感染的Vero细胞,以及牛蒡苷元作用的感染细胞在转录组水平上的差异;采用ELISA和荧光染色法检测细胞凋亡水平。结果:牛蒡苷元具有抑制细胞内ZIKV复制的活性,其抗病毒效果在一定浓度范围内呈量效关系,IC_(50)为(1.95±0.32)μg/mL。与ZIKV感染细胞相比,牛蒡苷元处理后的感染细胞共有4 612个基因表达发生显著改变,差异表达基因富集于凋亡相关信号通路,如Hippo和P53信号通路。ZIKV感染后表达上调而牛蒡苷元作用后表达下调(先上调后下调)的基因共376个。在前10个先上调后下调基因中,5个凋亡相关基因的mRNA表达水平验证了RNA-seq结果。细胞凋亡检测结果表明,牛蒡苷元降低了由ZIKV感染诱导的细胞凋亡水平。结论:牛蒡苷元可能通过调节宿主基因表达水平,抑制ZIKV感染诱导的细胞凋亡,发挥体外抗ZIKV活性。Objective:To explore the efficacy and mechanism of arctigenin(ATG)against Zika virus(ZIKV).Methods:The antiviral activities and action pattern of ATG against ZIKV were evaluated on African green monkey kidney cells(Vero)by real-time quantitative PCR(qRTPCR)and CPE assays.RNA sequencing(RNA-seq)was used to analyze the transcriptional differences among the normal Vero cells,the ZIKV-infected cells,and the infected cells with ATG treatment.Apoptosis was detected by ELISA and fluorescence staining.Results:ATG could inhibit the replication of ZIKV in cells,and its inhibitory effect was in a dose-dependent manner within a certain concentration range with IC_(50) of(1.95±0.32)μg/mL.Compared with ZIKA-infected cells,the infected cells with ATG treatment showed 4612 genes with significantly changed expression,and the differentially expressed genes(DEGs)were concentrated in apoptosis-related signaling pathways,such as Hippo and p53 signaling pathways.The expression of 376 genes was up-regulated after ZIKV infection and down-regulated after ATG treatment(referred to as up-to-down genes).Among the top 10 up-to-down genes,the mRNA expression levels of five apoptosis-related genes confirmed the results of RNA sequencing.The apoptosis detection results showed that ATG reduced cell apoptosis induced by ZIKV infection.Conclusion:ATG exerts in vitro anti-ZIKV activities by regulating host gene expression and inhibiting apoptosis induced by ZIKV infection.

关 键 词：牛蒡苷元 寨卡病毒 转录组测序 细胞凋亡 

分 类 号：R285[医药卫生—中药学]