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目的研究柯萨奇病毒(Coxsackievirus,CVB3)感染ECV304细胞后,血管细胞黏附分子-1(vascular cell adhesionmolecule,VCAM-1)的表达变化及海洋放线菌素X2对CVB3感染ECV304细胞VCAM-1表达的影响。方法采用MTT法检测不同浓度的海洋放线菌素X2作用病毒后的抑制情况;采用RT-PCR和FCM分别测定CVB3感染的ECV304细胞在海洋放线菌素X2作用前后不同时间点的VCAM-1的mRNA和蛋白的表达水平。结果海洋放线菌素X2在病毒吸附前对CVB3的SI为63.88。正常状态下ECV304细胞基本无VCAM-1 mRNA表达,CVB3感染ECV304细胞促进VCAM-1 mRNA表达,感染12 h达到最高峰,6～54 h时VCAM-1 mRNA水平,与细胞对照组相比CVB3感染组差异有统计学意义(P<0.05)。CVB3感染ECV304细胞后,VCAM-1蛋白表达在12～48 h显著高于正常ECV304细胞对照组(P<0.05)。海洋放线菌素X2干预后,于12～54 h降低ECV304细胞VCAM-1 mRNA的水平,12～24 h降低VCAM-1蛋白的表达,与CVB3感染组相比差异有统计学意义(P<0.05)。结论海洋放线菌素X2在病毒吸附前对CVB3具有抗病毒作用,并可下调CVB3诱导的ECV304细胞VCAM-1的mRNA水平和蛋白的表达。
Objective To investigate the expression of vascular cell adhesion molecule-1 (VCAM-1) in Coxsackievirus (CVB3) -induced ECV304 cells and the effect of actinomycin X2 on the expression of VCAM-1 in CVB3-infected ECV304 cells Impact. Methods MTT assay was used to detect the inhibitory effect of actinomycin-X2 at different concentrations. The levels of VCAM-1 in CVB3-infected ECV304 cells before and after actinomycin-X2 treatment were measured by RT-PCR and FCM respectively MRNA and protein expression levels. Results The marine actinomycin X2 had an SI of CVB3 of 63.88 before virus adsorption. ECV304 cells in normal condition showed no VCAM-1 mRNA expression, CVB3-infected ECV304 cells promoted VCAM-1 mRNA expression, peaked at 12 h, VCAM-1 mRNA level at 6 ~ 54 h, CVB3 infection The difference was statistically significant (P <0.05). The expression of VCAM-1 protein in CVB3-infected ECV304 cells was significantly higher than that in normal ECV304 cells (P <0.05) at 12-48 h. After actinomycin-X2 treatment, the level of VCAM-1 mRNA in ECV304 cells was reduced from 12 to 54 h, and the expression of VCAM-1 protein was decreased from 12 to 24 h, which was significantly different from that of CVB3-infected group (P < 0.05). Conclusion Actinomycin X2 has antiviral activity against CVB3 before virus adsorption and down-regulates the expression of VCAM-1 mRNA and protein in CVB3-induced ECV304 cells.