海洋放线菌素X2对CVB3感染ECV304细胞VCAM-1表达的影响  

Effects of marine actinomycin X2 on VCAM-1 expression in CVB3-infected ECV304 cells

作　　者：戴国奎 吴莎[2] 施珊珊[2] 孙毅凡[2] 江振友[2] 周琳 

机构地区：[1]广州市干部疗养院检验科,510530 [2]暨南大学医学院微生物学与免疫学系,广州510632 [3]广东省结核病控制中心,广州510630

出　　处：《免疫学杂志》2012年第7期589-594,共6页Immunological Journal

基　　金：科技部十二五重大科技专项(2012ZX10004903);国家自然科学基金资助课题(30973693)

摘　　要：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。To investigate antivirus affects of marine actinomycin X2 against Coxsackieviru （CVB3）, we detected the expression and secretion of vascular cell adhesion molecule （VCAM-1） in CVB3-infected ECV304 cells, and studied the effects of marine actinomycin X2 on the expression of VCAM-1 in CVB3-infected ECV304 cells. By adopting MrI^F methods, we detected the inhibitory action of virus by using different concentrations of marine actinomycin X2 and study the efficacy of viral adsorption. Cytoplasmic RNA from ECV304 cells was extracted by the Trizol method and mRNA levels of VCAM-1 were assayed by reverse transcript-polymerase chain reaction （RT- PCR）. Flow cytometer was used to detect ICAM-1 expression on the infected ECV304 in different time courses respectively. We found that marine actinomycin X2 could inhibit virus before absorption of CVB3 into the normal ceils and the SI was 63.88. The normal ECV304 did not express VCAM-1 mRNA in all 54 hours, but in the infected ECV304 cells, the expression of VCAM-1 mRNA was increased in 6-54 hours among which the VCAM-1 mRNA reached a peak at 12 hours point post-infection. The infection of CVB3 increased VCAM-1 protein expression on the surface of ECV304 at 12-48 hours. In CVB3＋X2 group, the expression of VCAM-1 mRNA was lower than that of CVB3 group at 12-54 hours （P〈0.05）, and the expression of VCAM-1 protein was lower than that of CVB3 group （P〈 0.05） at 12-24 hours. In conclusion, marine actinomycin X2 has anti-viral function, and has protective effects on the injured ECV304 exposed to CVB3.

关 键 词：海洋放线菌 柯萨奇病毒 血管细胞黏附分子 

分 类 号：R285.5[医药卫生—中药学]