摘要
为研究羧甲基茯苓多糖(CMP)对肠道病毒71型(EV71)的抗病毒活性,并探讨其作用机制,采用CCK-8法检测CMP对人恶性胚胎横纹肌瘤细胞(RD细胞)增殖的影响;采用致细胞病变效应(CPE)法观察CMP对EV71感染引起的CPE的抑制作用;采用致半数细胞病理改变的病毒感染剂量(TCID50)法检测CMP对EV71感染性病毒颗粒的抑制水平;采用RT-qPCR法检测EV71结构蛋白VP1的表达;采用Western blotting法分析VP1、P62、LC3、Caspase-3、cleaved Caspase-3蛋白的表达。结果表明:不同质量浓度(0、250、500、1000μg/mL)CMP对RD细胞无明显的细胞毒性;与对照组相比,CMP在无毒性浓度范围内可有效抑制EV71的复制,且能够呈剂量依赖性地降低病毒感染引起的细胞病变效应;CMP可呈剂量依赖性地降低EV71病毒结构蛋白VP1的表达和子代感染性病毒颗粒的产生;CMP可下调EV71病毒感染的RD细胞中的LC3-Ⅱ、cleaved Caspase-3的表达。
To investigate the antiviral activity and potential mechanism of carboxymethylpachyman(CMP)against enterovirus 71(EV71),the effect of CMP on the proliferation of RD cells was detected by CCK-8 method.The cytopathic effect(CPE)method was used to observe the inhibitory effect of CMP on the CPE caused by EV71 infection.The median tissue culture infective dose(TCID50)method was used to detect the inhibitory level of CMP on EV71 infectious virus particles.The expression of EV71 VP1 protein was detected by RT-qPCR.The expressions of VP1,P62,LC3,Caspase-3 and cleaved Caspase-3 protein were analyzed by Western blotting.The results showed that different mass concentrations(0,250,500,1000μg/mL)of CMP had no significant cytotoxicity on RD cells.Compared with the control group,CMP effectively inhibited the replication of EV71 in the range of non-toxic concentrations of EV71 replication and dosedependently reduced the cytopathic effect caused by viral infection.CMP dose-dependently reduced the expression of EV71 viral structural protein VP1 and decreased the production of infectious viral particles in the offspring.CMP down-regulated the expression of LC3-II and cleaved Caspase-3 after EV71 virus infection of RD cells.
作者
曾露芝
吴春晨
向蒙
彭逸斯
王哲
汪启明
刘实
彭国平
ZENG Luzhi;WU Chunchen;XIANG Meng;PENG Yisi;WANG Zhe;WANG Qiming;LIU Shi;PENG Guoping(Hunan Engineering Laboratory of Standardized Cultivation and Comprehensive Utilization of Indigenous Medicinal Plants,Hunan Agricultural University,Changsha 410128,China;Department of Clinical Laboratory,Hubei Maternal and ChildHealth Hospital,Wuhan 430070,China;State Key Laboratory of Virology,Wuhan University,Wuhan,Hubei 430072,China)
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2024年第5期104-111,共8页
Journal of Hunan Agricultural University(Natural Sciences)
基金
湖湘高层次人才聚集工程项目(2021RC5006)
湖北省自然科学基金项目(2022CFB564)。
关键词
羧甲基茯苓多糖
肠道病毒71型
抗病毒活性
carboxymethyl porphyra polysaccharide
enterovirus 71
antiviral activity
作者简介
曾露芝(1997—),女,湖南益阳人,硕士研究生,主要从事中草药资源综合开发与中草药生物技术研究,570874673@qq.com;通信作者:彭国平,博士,副教授,主要从事中药资源开发与生物技术、有效成分分离纯化与功能开发、中草药规范化栽培与加工研究,pgphh@163.com。
