小儿青翘颗粒抗肠道病毒EV71的药效作用及机制研究  

The antiviral effect of Xiaoer Qingqiao granule against enterovirus 71

作　　者：王荣花 苏勤 郑志慧 王书源 毛旭华 卫海琳 王雯蕾 陈晓[4] 张评浒 WANG Ronghua;SU Qin;ZHENG Zhihui;WANG Shuyuan;MAO Xuhua;WEI Hailin;WANG Wenlei;CHEN Xiao;ZHANG Pinghu(Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases/College of Medicine(Institute of Translational Medicine),Yangzhou University,Yangzhou 225009,China;YifanPharmaceuticalCompanyLimited,Hangzhou311300,China;Department of Clinical Laboratory,the Affiliated Yiaing Clinical College of Medicine of Yangzhou University,Yixing 214200,China;Department of Pediatric,the Af filiated Hospital of Yangzhou University,Yangzhou 22509,China)

机构地区：[1]扬州大学医学院(转化医学研究院)/江苏省中西医结合老年病防治重点实验室,江苏扬州225009 [2]亿帆医药股份有限公司,杭州311300 [3]扬州大学医学院宜兴临床学院检验科,江苏宜兴214200 [4]扬州大学附属医院儿科,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2022年第4期75-82,共8页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家重大新药创制科技专项(2019ZX09731001);扬州大学高端创新人才资助项目(137011384);扬州大学大学生创新创业训练计划项目(X20200766)。

摘　　要：肠道病毒71型(enterovirus 71, EV71)是引起手足口病(hand, foot and mouth disease, HFMD)的主要病原体,在全球周期性暴发流行,至今尚无特效药物。临床研究显示中成药在治疗小儿手足口病上具有一定疗效。小儿青翘颗粒(XEQQ)是由山银花、连翘、葛根、大青叶等多味中药组成的中药方剂,临床研究发现其对EV71引起的手足口病症具有一定疗效,但其是否具有直接抗EV71作用及其机制尚未见报道。以EV71感染Vero细胞为模型,采用CPE、qRT-PCR、Western blot等方法探究XEQQ抗EV71的作用及其潜在机制。结果表明:XEQQ在0.125~4 mg·mL^(-1)剂量范围内对Vero细胞无明显细胞毒性,在上述剂量范围内能剂量依赖性地抑制EV71病毒的复制及其病毒RNA与蛋白质的转录表达,其抑制EV71病毒复制的半数有效浓度IC为76μg·mL^(-1)。此外,XEQQ还能有效抑制EV71感染所致炎症因子IL-6、TNF-α和IL-1β mRNA转录表达。XEQQ可能通过促进自噬标志物LC3B-II和P62聚集阻断EV71诱导自噬促进自身复制的机制而最终抑制EV71的病毒复制。该研究为XEQQ临床治疗手足口病提供了理论依据。Enterovirus 71(EV71) is the main pathogen of hand, foot and mouth disease(HFMD), which was pandemic all over the world and absent of specific antiviral drugs. Clinical researches indicate that Chinese traditional medicine exhibited a good therapeutic effect on HFMD by reducing clinical symptoms. Xiaoer Qingqiao(XEQQ) granules, which is composed of Flos Lonicerae, Forsythia suspensa, Pueraria lobata, Radix Isatidis and other Chinese traditional medicines, showed a good therapeutic effect on HFMD. However, the detailed antiviral effect and underlying mechanism remains unclear. In this study, the antiviral effect and potential mechanism of XEQQ against EV71 were for the first time evaluated with CPE, qRT-PCR and Western blot methods. Our results showed that 0.125-4 mg·mL^(-1)of XEQQ had no obvious cytotoxicity on Vero cells, but could dose-dependently inhibit the replication of EV71 by suppressing the transcription of viral VP1RNA and the expression of VP1 protein,with 76μg·mL^(-1)of IC.Furthermore,XEQQ could effectively inhibit the mRNA levels of inflammatory cytokines such as IL-6,TNF-α and IL-1β caused by EV71 infection.Mechanistic researches indicated that XEQQ might block EV71-induced autophagy by promoting the aggregation of autophagy markers LC3B-II and P62 to inhibit the replication of EV71.Our results provide a theoretical basis for the clinical treatment of hand,foot and mouth disease with XEQQ.

关 键 词：小儿青翘颗粒 手足口病 EV71 炎症因子 IL-6 自噬 

分 类 号：R285[医药卫生—中药学]