大黄素体外抗柯萨奇病毒B_4的实验研究

Antiviral Effect of Emodin on Coxsakievirus B_4 in vitro

为研究大黄素的体外抗柯萨奇病毒B4（CVB4）作用,以病毒唑为阳性对照药,用MTT法从药物抑制病毒增殖、直接杀灭和抗吸附3种方式评价了大黄素抗CVB4效果,并用实时定量PCR检测了大黄素对CVB4基因mRNA表达的影响,用空斑法测定了大黄素对病毒感染细胞不同时间空斑形成率的影响.结果表明：大黄素不能直接杀灭病毒或阻断病毒吸附细胞,但能显著抑制病毒在细胞内的生物合成,其IC50=（14.10±0.74）μM,TI=（4.35±0.60）,能显著降低细胞内CVB4基因mRNA表达（p〈0.05）,其抑制作用发生在病毒进入细胞0-4 h.故大黄素主要通过抑制CVB4穿入细胞后复制环节发挥抗病毒作用,并具有时间依赖性和剂量依赖性.

To investigate the antiviral effect of emodin on Coxsakievirus B4,ribavirin was used as the positive control and MTT assay was used to evaluate its anti-CVB4 effect through 3 manners including propagation inhibition,direct inactivation and anti-absorption. Then real-time PCR were performed to determine the effect of emodin on the mRNA expression of CVB4 and plaque forming assay were applied to detect the effect of emodin on the plaque formed rate after virus infection at different time. The results indicated that emodin could not directly inactivate the viruses or block their absorption to cells,but it may act as a biological synthesis inhibitor. The IC50 was detected as（ 14. 10 ± 0. 74） μM and TI were detected as（ 4. 35 ± 0. 6）. Emodin could significantly reduce the mRNA expression of CVB4 gene and may act its inhibitory effect during 0 - 4 h postinfection（ p〈0. 05）. So emodin may act its antiviral activity of inhibiting CVB4 replication after virus penetration in a concentration-and time-dependent manner.