人巨细胞病毒和糖基化终产物共同作用对血管内皮细胞的影响

Co-effects of human cytomegalovirus infection and advanced glycation end products on vascular endothelial cells

目的:观察人巨细胞病毒(human cytomegalovirus,HCMV)和糖基化终产物(advanced glycation endproducts,AGEs)共同作用对血管内皮细胞(vein endothelial cells,ECV)氧化应激的影响及对糖基化终产物受体(the receptor for advanced glycation end products,RAGE)表达的影响,明确HCMV和AGEs在致内皮细胞损伤作用中是否具有协同效应,探讨HCMV和AGEs致动脉粥样硬化(AS)发生和发展的可能机制。方法:用HCMV和AGEs分别及共同作用于ECV,将实验对象分为:对照组、HCMV组、牛血清白蛋白(BSA)组、AGEs组、AGEs+HCMV组5组;用激光共聚焦显微镜检测细胞内活性氧(ROS)的改变;采用RT-PCR方法检测血管内皮细胞RAGEmRNA的表达。结果:HCMV感染ECV后,对照组荧光强度和BSA组荧光强度均较弱,两组间比较差异无统计学意义(P>0.05);AGEs组和HCMV组荧光强度强于对照组(P<0.01);AGEs+HCMV组荧光强度高于AGEs组和HCMV组(P<0.05),两者联合作用存在协同效应。对照组和BSA组ECV细胞RAGEmRNA均有低水平表达,两组之间比较无差异(P>0.05);AGEs组和HCMV感染组RAGEmRNA表达量高于对照组(P<0.01);AGEs+HCMV组表达量高于AGEs组和HCMV组(P<0.01)。在相同病毒滴度感染的情况下,随AGEs浓度增加RAGEmRNA的表达增高,呈浓度依赖性;在相同AGEs浓度作用时,随HCMV感染滴度增加RAGEmRNA的表达水平升高,呈滴度依赖性。各组之间比较差异有统计学意义(P<0.05)。结论:HCMV和AGEs均能够增强血管内皮细胞氧化应激,促进RAGEmRNA的表达,两者共同作用时呈协同效应。HCMV和AGEs有可能通过协同增强氧化应激、进一步上调RAGEmRNA的表达,介导血管内皮细胞的炎症反应,促进动脉粥样硬化的发生、发展。

Objective： To study the mechanisms of human cytomegalovirus（HCMV） infection in atherosclerosis by investigating changes of reactive oxygen species （ROS） and sequential changes of the receptor of advanced glycation end products （RAGE） in vascular endothelial cells with human cytomegalovirus infection. Methods： Vascular endothelial cells were cultured and then infected with HCMV, then changes of reactive oxygen species were identified by confocal microscopy; Expression of RAGEmRNA was determined by the reverse transcriptase polymerase chain reaction. Results： Fluorescence intensity was determined at a low level in the control group and the BSA group, with no significant difference （ P 〉 0.05）. However, it was obviously higher in the AGEs group and the HCMV group than in the control group （P 〈 0.01） and it was further increased in the HCMV combined with AGEs group, which was significantly higher than that in the AGEs and the HCMV groups （ P 〈 0.01 ）. RAGEmRNA was expressed at a low level in the control group and the BSA group, with no significant difference （ P 〉 0.05）. However, it was obviously higher in the HCMV group and the AGEs group than in the control group （ P 〈 0.01 ）, and was further increased in the HCMV combined with AGEs group, which was significantly higher than that in the AGEs group and the HCMV group （ P 〈 0.01 ）. RAGE mRNA expression gradually increased by an increase of AGEs concentration with the same trites of HCMV in a concentration-dependent manner and was also gradually enhanced by an increase of HCMV trite with the same concentration of AGEs in a trite-dependent manner. Conclusions： Both HCMV infection and AGEs can enhance the oxidative stress and the expression of RAGEmRNA in vascular endothelial cells in a co-effects manner. Both HCMV and AGEs have an effect on the injury of vascular endothelial cells by enhancing the oxidative stress and up-regulating the RAGE expression on endothelial cells, which further facilitates the occurrence and development of atherosclerosis.