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人巨细胞病毒感染致血管内皮细胞损伤机制的研究.doc

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人巨细胞病毒感染致血管内皮细胞损伤机制的研究 作者:王凤 作者单位:实验畸形学教育部重点实验室山东大学医学院微生物学教研室, 济南 250012 【摘要】 目的 观察人巨细胞病毒(HCMV)感染对人血管内皮细胞氧化应激的影响及对血管细胞粘附分子1(VCAM1)和糖基化终产物受体(RAGE)时序性表达的影响,研究HCMV感染致动脉粥样硬化(AS)的作用机制。方法 用HCMV感染血管内皮细胞,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方法检测不同感染时段细胞VCAM1及RAGEmRNA的表达。结果 HCMV感染血管内皮细胞后,HCMV组荧光强度显著高于对照组(P<0.01)。感染0?h时,VCAM1mRNA有基础水平的表达,4?h开始升高,8?h时达高峰,12?h开始回落,24?h回落更明显,与0?h比较,均有统计学意义(P<0.01),48?h接近0?h表达水平(P>0.05)。感染0?h时,RAGEmRNA有基础水平的表达,4?h开始升高,8?h时表达水平明显升高,随感染时间延长,表达量逐渐增高,感染24?h时达高峰,48?h开始回落,仍维持在较高的水平,各时段与0?h比较,均有统计学意义(P<0.01)。结论 HCMV感染血管内皮细胞后能够增强氧化应激,促进VCAM1及RAGE的mRNA表达,并且呈时间依赖性。HCMV有可能通过增强氧化应激、上调VCAM1及RAGE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。 【关键词】 巨细胞病毒 脐静脉 活性氧 血管细胞粘附分子1 糖基化终产物 受体 WANG Feng, WU Jianmin, ZHOU Yabin, LI Shuying, CHENG Yizhe, QI Mei, TANG Wei, WANG Hong, LIU Juan, YU Han (Key Laboratory of Experimental Teratology, Ministry of Education, China; Department of Microbiology, School of Medicine, Shandong University, Jinan 250012, China) Abstract: Objective To explore the mechanisms of cytomegalovirus infection by investigating the changes of reactive oxygen species (ROS) and sequential changes of the vascular cell adhesion molecule1 (VCAM1) and the receptors for advanced glycation end products (RAGE) in vascular endothelial cells with human cytomegalovirus(HCMV) infection. Methods Vascular endothelial cells were cultured and then infected by HCMV, then changes of reactive oxygen species were identified by confocal microscopy. Expressions of VCAM1 and RAGE mRNA were determined by reverse transcriptase polymerase chain reaction. Results Fluorescence intensity was determined at a low level in the control group and was significantly increased in the HCMV group (P<0.01). VCAM1 mRNA was expressed at a low level at 0 hour, began to increase at 4 hours after HCMV infection and reached a peak at 8 hours. After 12 hours, it began to decrease and significantly decreased at 24 hours, but
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