木犀草素对肠道病毒71型感染细胞的影响

Luteolin inhibits enterovirus 71 induced cytopathic effects in vitro

目的:探讨木犀草素体外抗肠道病毒71型(enterovirus 71,EV71)感染的机制。方法:分别用0、25、50、100、200、400μg/m L木犀草素处理人横纹肌肉瘤(RD)细胞,48 h后检测细胞存活率;另将RD细胞分为对照组(常规培养)、木犀草素组(25、50、100μg/m L)、EV71感染组和木犀草素(25、50、100μg/m L)+EV71感染组,于病毒感染48 h后,MTT法检测各组细胞存活率,流式细胞术检测细胞凋亡,蛋白质印迹法检测Caspase-3表达,ELISA法检测细胞TNF-α,IL-β,IL-6和IL-10浓度。结果:与0μg/m L相比,25、50、100μg/m L木犀草素对RD细胞存活率无明显影响(P>0.05);25、50和100μg/m L木犀草素+EV71感染组RD细胞存活率均明显高于EV71感染组(P均<0.05);与EV71感染组相比,木犀草素+EV71感染组细胞凋亡率和Caspase-3表达明显降低,TNF-α、IL-1β、IL-6和IL-10分泌明显减少(P均<0.05)。结论:木犀草素可能通过抑制细胞凋亡和减少促炎症因子分泌,抑制EV71对细胞的感染。

Objective：To investigate the mechanism of inhibition of enterovirus 71 （EV71）induced cy-topathic effects by luteolin in vitro.Methods：Rhabdomyosarcoma（RD）cells were treated with 0,25,50, 100,200,400 μg/mL luteolin for 48 h,then cells viability was determined by MTT.Meanwhile,RD cells were divided into control group （DMEM treatment）,luteolin group （25,50,100 μg/mL luteolin treat-ment）,EV71 infection group and luteolin（25,50,100 g/mL）＋EV71 infection group.The cell viability was assessed by MTT assay.Apoptosis of RD cells was detected by flow cytometry.The expression of Caspase-3 was detected by Western blotting.In addition,the concentration of pro-inflammatory cytokines were detected by ELISA.Results：Compared with 0 μg/mL luteolin,25,50 or 100 μg/mL luteolin had no significant effect on RD cell viability（P 〉0.05）.The survival rates of RD cells in 25,50 or 100 μg/mL luteolin ＋EV71 group were significantly higher than EV71 group（all P 〈0.05）.Compared with EV71 group,the apoptosis rate,Caspase-3 expression and the concentration of TNF-α,IL-1β,IL-6 and IL-10 were significantly decreased in luteolin ＋EV71 group（all P 〈0.05）.Conclusion：Luteolin inhibits EV71 infection induced cytopathic effects and these protection may be resulted from the inhibition of apoptosis and inflammatory cytokine secretion.