登革病毒感染对人血管内皮细胞血管细胞黏附分子-1表达的影响  

Effect of Dengue virus infection on expression of vascular cell adhesion molecule-1 in human vascular endothelial cells

作　　者：肖瑞[1] 江振友[2] 张倩 

机构地区：[1]广州医学院临床技能实验中心,广州510182 [2]暨南大学医学院微生物学与免疫学教研室,广州510632 [3]安徽省芜湖市疾病预防与控制中心,芜湖241000

出　　处：《第二军医大学学报》2008年第7期750-755,共6页Academic Journal of Second Military Medical University

基　　金：广东省医学科研基金(A2005340)~~

摘　　要：目的:研究登革病毒2型(DV2)体外感染对人脐静脉内皮细胞(HUVEC)表达血管细胞黏附分子-1(VCAM-1)的影响。方法:原代分离、纯化、培养人HUVEC细胞,用生长良好的2 ～3代细胞进行实验。分别用病毒感染复数(MOI)为1、2、4、8、 16的DV2病毒液感染HUVEC,阴性对照组直接以RPMI 1640培养,应用细胞计数试剂-8(CCK-8)法测定DV2感染前和感染后 6、12、24、48、72和96 h的细胞活性。另以MOI =2的DV2病毒液感染细胞,阴性对照组直接以RPMI 1640培养,于感染后6、12、 24、48、72、96 h分别收集病毒感染的HUVEC,采用流式细胞术测定细胞表面VCAM-1表达水平;采用RT-PCR法检测细胞中 VCAM-1 mRNA转录水平。结果:当病毒MOI =2时对细胞活力的影响与对照组相比差异无统计学意义。DV2感染可以促进 HUVECs中VCAM-1 mRNA的转录,96 h内均有增加,与对照组相比有显著性差异(P<0.05)。其中,正常状态下HUVEC基本无VCAM-1 mRNA转录,感染12 h达到最高峰,12 ～48 h表达量显著高于其他时间(P<0.05)。DV2感染HUVEC后,VCAM-1蛋白表达在12 ～72 h显著升高(P<0.05),与对照组相比差异有统计学意义(P<0.05)。结论:DV2感染上调HUVEC 的VCAM-1 mRNA转录和蛋白表达。这可能是DV2感染诱发患者血管通透性升高和血浆渗漏的重要分子机制之一。Objective：To investigate the effects of Dengue virus 2 （DV2） on the expression and secretion of vascular cell adhesion molecule-1 （VCAM-1）in human umbilical vein endothelial cells （HUVECs）. Methods： HUVECs were primarily isolated, purified and cultured. Cells of 2-3 generations were infected with DV2 with different MOI values （1,2,4,8 and 16）. HUVECs cultured with RPMI1640 medium were taken as negative control. The viability of HUVECs was assessed by CCK-8 assay before and at different time points after DV2 infection（6 h, 12 h, 24 h,48 h, 72 h and 96 h）. Flow cytometry was used to detect the VCAM-1 expression in HUVECs. The expression of cytoplasmic VCAM-1 mRNA was assayed by reverse transcription-polymerase chain reaction （RT-PCR）. Results：When the MOI value was 2,DV2 had no significant influence on cell viability compared with the control group. DV2 infection significantly promoted the transcription of VCAM-1 mRNA compared with the control group（P〈0.05）. The expression of VCAM-1 mRNA peaked at 12 h after infection and remained at a higher level within 96 h（P〈0.05）. The expression at 12-48 h after infection was significantly higher than that at other time points（P〈0. 05）. HUVECs hardly had VCAM-1 mRNA expression under normal condition. The expression of VCAM-1 protein was significantly increased at 12-72 h after infection compared with the control group （P〈0.05）, Conclusion：DV2 can increase the expression of VCAM-1 mRNA in HUVECs, which might be one of the important mechanisms for elevated vascular permeability and plasma leakage after DVeinfection.

关 键 词：登革病毒 人脐静脉内皮细胞 血管细胞黏附分子-1 

分 类 号：R373.33[医药卫生—病原生物学]