Objective To investigate the effect of daphnetin on RD cells infected with EV71. Methods The experiment was set up in 4 groups, normal group(RD cells), virus control group(RD cells infected with EV71), daphnetin group(RD cells infected with EV71 were treated with different concentrations of daphnetin)and positive drug group(RD cells infected with EV71 and treated with RVB). CCK8 assay, cytopathic effect(CPE)and real-time RT-PCR were used to determine the effect of daphnetin against EV71 virus at different stages of viral infection. Results The 50％cytotoxicity(TC50)of daphnetin on RD cells was 33.47 μg/m L, and the maximum nontoxic concentration(TC0)was 15μg/m L. The daphnetin of 3.75-15 μg/m L was used during EV71 infection and late-infection. In particular, 1 daphnetin of 2.5-15 μg/m L had a relatively good therapeutic effect, and not significantly affected the cells compared with normal cell group(P＞ 0.05). Daphnetin of 15 and 10 μg/m L administered after EV71 infection significantly inhibited the virus replication, and daphnetin of 15, 10 and 7, 5 μg/m L administered at the time of EV71 infection also significantly inhibited the virus replication. Conclusion Daphnetin can effectively inhibit the replication of EV71 and expected to provide experimental evidence for the development of new anti-EV71 drugs.%目的 探讨瑞香素对肠道病毒71型(EV71)感染人恶性胚胎横纹肌肉瘤(RD)细胞的影响.方法 实验设定为4组:正常细胞组指不加药物及EV71干预RD细胞;病毒对照组指感染EV71的RD细胞;瑞香素组指不同浓度(根据瑞香素最大无毒浓度设定瑞香素作用浓度为15、12.5、10、7.5、5、3.75、2.5、1.875、1.25、1μg/m L)的瑞香素作用于感染EV71的RD细胞;阳性药物组是指50μg/m L利巴韦林(ribavirin, RVB)作用于感染EV71的RD细胞.运用CCK8测定法、细胞病变效应(CPE)、RT-PCR方法测定瑞香素在病毒感染不同阶段的抗EV71病毒作用.结果 瑞香素对RD细胞的50％细胞毒性浓度(TC50)为33.47μg/m L, 最大无毒浓度(TC0)为15μg/m L.浓度为3.75～15μg/m L瑞香素在EV71感染期和感染后期对EV71均有较好的抑制效果, 其中12.5、15μg/m L瑞香素的治疗效果相对较佳, 其病毒抑制率与正常细胞组比较差异无统计学意义(P＞ 0.05).浓度为15、10μg/m L的瑞香素在EV71感染细胞后再用药处理, 能有效降低病毒载量;而以浓度为15、10、7.5μg/m L瑞香素与EV71病毒混合作用于细胞, 同样具有降低病毒载量的作用, 均对EV71的复制有较好的抑制效果, 差异有统计学意义(P ＜0.05).结论 瑞香素能有效抑制EV71的复制, 有望为开发新的抗EV71药物提供实验依据.
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