穿心莲内酯体外抗肠道病毒71型和柯萨奇病毒A16型的作用  

Effect of andrographolide on enterovirus 71 and coxsackievirus A16 in vitro

作　　者：纪洵敏 梁宇恒 何炽明 肖微 彭晓放 谭丽容 曾宏强 唐颜 柯昌文[2] 张磊 JI Xunmin;LIANG Yuheng;HE Chiming;XIAO Wei;PENG Xiaofang;TAN Lirong;ZENG Hongqiang;TANG Yan;KE Changwen;ZHANG Lei(Guangdong Provincial Institute of Biological Products and Materia,Guangzhou,Guangdong 510440,China;Guangdong Provincial Center Disease Control and Prevention,Guangdong Provincial Key Laboratory of Pathogen Detection for Emerging Infectious Disease Response,Guangzhou,Guangdong 510430,China)

机构地区：[1]广东省生物制品与药物研究所,广东广州510440 [2]广东省疾病预防控制中心,广东省新发传染病应急检测技术研究与应用重点实验室,广东广州511430

出　　处：《中国热带医学》2025年第1期62-68,共7页China Tropical Medicine

基　　金：“广东特支计划”卫生健康平台科技创新人才项目(No.0620220104);广东省科技计划项目(No.2023B1212010010);广东省中医药局科研项目(No.20211054,No.20222024);广东省医学科学技术研究基金项目(No.C2022015,No.B2023031,No.B2023032,No.A2024276,No.A2024268)。

摘　　要：目的观察穿心莲内酯体外抗肠道病毒71型(enteroviruses 71,EV71)和柯萨奇病毒A16型(coxsackievirus A16,CA16)的作用效果。方法采用细胞病变(cytopathic effect,CPE)法和噻唑蓝(thiazolyl blue tetrazolium bromide,MTT)法测定穿心莲内酯对人恶性胚胎横纹肌瘤细胞(human rhabdomyosarcoma cells,RD)的最大无毒剂量和穿心莲内酯对感染细胞后的EV71和CA16体外抑制作用;通过荧光定量PCR法检测穿心莲内酯溶液对EV71和CA16中VP1基因,RD细胞中3种促炎因子IL-1β、IL-6和TNF-α在基因水平的变化。结果MTT结果显示穿心莲内酯对RD细胞的最大无毒剂量为78μg/mL;手足口病毒感染细胞,随后分别经78.00、39.00、19.50、9.75、4.88、2.44μg/mL的穿心莲内酯溶液处理,感染EV71病毒的细胞存活率增加至(82.41±1.76)%、(79.54±2.91)%、(81.02±1.99)%、(71.81±2.26)%、(52.87±1.51)%和(50.41±0.93)%,感染CA16病毒的细胞存活率增加至(81.00±0.64)%、(79.72±1.38)%、(61.59±3.47)%、(53.37±0.53)%、(52.41±1.37)%和(43.69±0.40)%,表明穿心莲内酯能明显降低EV71和CA16引起的细胞病变效应;手足口病毒感染后,用78.00、39.00、19.50、9.75μg/mL的穿心莲内酯溶液给药,显示EV71和CA16中VP1,RD细胞中促炎因子IL-1β、IL-6和TNF-α表达量与病毒对照组相比均有明显降低,结果表明手足口病毒感染后,用穿心莲内酯溶液给药,对手足口病毒的VP1基因表达产生明显抑制作用,且能降低IL-1β、TNF-α、IL-6在mRNA水平的表达。结论穿心莲内酯具有明显的体外抗EV71和CA16病毒的效果,其可能通过抑制促炎因子IL-1β、TNF-α、IL-6的表达,从而发挥抗EV71和CA16病毒的作用。Objective To investigate the effect of andrographolide on enterovirus 71(EV71)and coxsackievirus A16(CA16)in vitro.Methods Cytopathic CPE assay and thiazolyl blue tetrazolium bromide(MTT)assay were used to determine the maximum non-toxic dose of andrographolide on RD cells and the inhibitory effect of andrographolide on EV71 and CA16 infection in vitro.The effects of andrographolide on the virus VP1 gene and cellular inflammatory factors(IL-1β,IL-6,and TNF-α)at gene levels were detected by quantitative fluorescence PCR.Results MTT results showed that the maximum non-toxic dose of andrographolide on RD cells was 78.00μg/mL.Upon infection with hand-foot-mouth disease viruses,cells treated with andrographolide solutions at concentrations of 78.00,39.00,19.50,9.75,4.88,and 2.44μg/mL showed increased survival rates to(82.41±1.76)%,(79.54±2.91)%,(81.02±1.99)%,(71.81±2.26)%,(52.87±1.51)%,and(50.41±0.93)%for EV71 and(81.00±0.64)%,(79.72±1.38)%,(61.59±3.47)%,(53.37±0.53)%,(52.41±1.37)%,and(43.69±0.40)%for CA16,respectively,indicating a significant reduction in the cytopathic effect caused by EV71 and CA16.When infected with hand-foot-mouth viruses and treated with andrographolide solutions at concentrations of 78.00,39.00,19.50,and 9.75μg/mL,the expression levels of VP1 in EV71 and CA16,along with pro-inflammatory factors IL-1β,IL-6,and TNF-αin RD cells,were significantly lower compared to the virus control group.These results indicated that after infection with hand-foot-mouth viruses,treatment with andrographolide solution significantly inhibits the expression of the VP1 gene and reduces the mRNA levels of IL-1β,IL-6,and TNF-α.Conclusions Andrographolide exhibits obvious in vitro antiviral effects against EV71 and,potentially through the inhibition of the expression of pro-inflammatory factors IL-1β,TNF-α,and IL-6,thereby exerting its antiviral effects.

关 键 词：穿心莲内酯 手足口病 肠道病毒71型 柯萨奇病毒A16型 抗病毒作用 

分 类 号：R512.5[医药卫生—内科学]