五肽对肠道病毒71型在人横纹肌肉瘤细胞和小鼠体内复制的抑制作用

Pentapeptides prevent enterovirus 71 proliferation in rhabdomyosarcoma cells and mice

肠道病毒71型(EV71)是手足口病(HFMD)主要的病原体之一。为研究小分子肽类化合物抑制EV71感染的作用,采用五肽分子作为干预分子,考察其对EV71在人横纹肌肉瘤(RD)细胞和乳鼠模型体内复制的影响。通过Western blotting、real-time PCR方法,检测EV71病毒蛋白和mRNA的表达水平,使用显微镜观察细胞病变效应。通过乳鼠体重变化和肠组织中EV71病毒VP1蛋白表达水平的变化来评价五肽体内抗病毒活性。结果表明,五肽分子P010157能够显著抑制RD细胞中EV71 VP1蛋白的表达,其质量浓度为100μg·mL-1时,EV71 vp1 mRNA水平降低了(92.0±6.3)%,EC50约为2.2μg·mL-1,且能有效抵抗病毒感染产生的细胞病变效应。通过MTS比色法测定P010157对RD细胞的细胞毒活性,结果显示其对细胞无明显毒性。体内实验结果还表明,P010157对感染EV71的乳鼠有保护作用(乳鼠无明显消瘦现象),并能抑制EV71在乳鼠体内的复制。本研究证实,P010157能有效抑制体外及体内的EV71的复制,是一种新型的对抗手足口病病毒EV71的化合物。

Enterovirus 71 （EV71） is the main causative agent of hand, foot, and mouth disease （HFMD）. This article presented the inhibitory activity of pentapeptides on the EV71 infection in rhabdomyosarcoma （RD） and suckling mice. The EV71 VP1 capsid protein expression levels and mRNA levels were analyzed by Western blotting and real-time PCR. The antiviral activity of pentapeptides in vivo was evaluated by weight changes and EV71 VP1 protein expression levels in intestines of suckling mice. Results revealed that the pentapeptide P010157 was able to inhibit EV71 replication in RD cells. After being incubated with the P010157 at a concentration of 100 μg·mL^-1 for 48 h, the level of EV71 vpl mRNA in RD cells decreased by （92.0±6.3）%. The estimated EC50 was 2.2 μg·mL^-1. P010157 was able to inhibit EV 71-induced cytopathic effect （CPE） in RD cells. The cytotoxic activity of the compound was evaluated against RD cells by MTS assay. The results showed that P010157 had no obvious toxicity. In addition, the treated mice with P010157 did not exhibit weight loss, as was observed in untreated mice. EV71 replication reduced significantly as revealed by Western blotting. These findings suggest that P010157 could prevent EV71 proliferation in vitro and in vivo. P0 l0157 is a novel compound for antiviral therapies against EV71, which merited further investigation.