| 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。 |
| 关键词: 登革病毒 人脐静脉内皮细胞 血管细胞黏附分子-1 |
| DOI：10.3724/SP.J.1008.2008.00750 |
| 投稿时间：2007-11-28修订日期：2008-03-17 |
| 基金项目:广东省医学科研基金(A2005340). |
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| Effect of Dengue virus infection on expression of vascular cell adhesion molecule-1 in human vascular endothelial cells |
| XIAO Rui1，JIANG Zhen-you2*，ZHANG Qian3 |
| (1.Clinical Skill Center,Guangzhou Medical College,Guangzhou 510182,China;2.Department of Microbiology and Immunology,Medical College of Ji’nan University,Guangzhou 510632;3.Wuhu Center for Disease Control and Prevention of Anhui Province,Wuhu 241000) |
| Abstract: |
| 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 DV2 infection. |
| Key words: dengue virus human umbilical vein endothelial cells vascular cell adhesion molecule-1 |
