摘要
研究背景人巨细胞病毒(HCMV)与动脉粥样硬化、移植性血管硬化和动脉再狭窄密切相关。已有研究证实人巨细胞病毒感染可诱发血管新生继而引起血管性疾病。因此,本文拟进一步探讨人巨细胞病毒诱导内皮细胞血管新生的分子学机制。方法分别于感染HCMV2、6、12和24h后收集血管内皮细胞EA.hy926,实时定量聚合酶链反应和Western blotting法检测SIRT1 mRNA及其蛋白质表达水平;通过激动剂Resveratrol或抑制剂SIRT1 siRNA孵育内皮细胞,观察病毒感染24h后其增殖、迁移和小管生成能力。结果与模拟感染组相比,HCMV感染组EA.hy926细胞SIRT1 mRNA表达无变化(F=1.395,P=0.304);而SIRT1蛋白表达水平呈逐渐递减趋势(F=23.927,P=0.000)。激动或抑制SIRT1蛋白表达后,HCMV感染组EA.hy926细胞迁移(P=0.008,0.003)和成管能力(P=0.012,0.008)下降或增强;而细胞增殖能力无变化(P=0.969,0.948)。结论 HCMV感染血管内皮细胞后能够促进细胞增殖、迁移和小管生成能力,其作用机制可能与抑制SIRT1蛋白表达有关。
Background atherosclerosis,transplanted vascular sclerosis and arterial restenosis.It has been proved that angiogenesis Human cytomegalovirus（HCMV） is closely related to diseases including induced by HCMV infection could result in vascular diseases.This article aims toEndothelial the mechanism of angiogenesis induced by HCMV infection in endothelial cells.Methods（EA.hy926 cells） were divided into HCMV infected group and mock infected group.The cells were investigatecells collected at 2,6,12 and 24 h after infection.Quantitative real-time polymerase chain reaction（qRT-PCR） and Western blotting were used to analyze SIRT1 mRNA and protein levels.Endothelial cells were incubated respectively by Resveratrol and SIRT1 small interference RNA（siRNA） for 2 h before infection.Twenty-four hours after infection,the proliferation,migration and tubule formation of cells were assessed by CCK-8,migration assay and tubule formation assay to detect the angiogenic response of endothelial cells.Results remained unchanged（F = 1.395,P = 0.304）,but the expression of SIRT1 protein decreased gradually（F = Compared to mock infected group,the expression of SIRT1 mRNA in HCMV infected group 23.927,P = 0.000）.Under the treatment of Resveratrol and SIRT1 siRNA,migration（P = 0.008,0.003） and tubule formation（P = 0.012,0.008） of endothelial cells increased or reduced.The proliferation,however,remained unchanged（P = 0.969,0.948）.Conclusion HCMV infection promotes proliferation,migration and tubule formation of endothelial cells,and its mechanism may be related to the supression of SIRT1.
出处
《中国现代神经疾病杂志》
CAS
2013年第1期36-42,共7页
Chinese Journal of Contemporary Neurology and Neurosurgery
基金
国家自然科学基金资助项目(项目编号:81271311)
国家自然科学基金资助项目(项目编号:81041020)
北京市自然科学基金资助项目(项目编号:7102040)
北京市自然科学基金资助项目(项目编号:7132060)
北京市卫生系统高层次卫生技术人才培养计划项目(项目编号:20113004)
北京市出国人员择优基金资助项目~~
关键词
巨细胞病毒
抗衰老酶
内皮细胞
动脉硬化
转染
细胞增殖
藜芦
RNA
小分子干扰
Cytomegalovirus
Sirtuins
Endothelial cells
Arteriosclerosis
Transfection
Cell proliferation
Veratrum nigrum
RNA
small interfering
