摘要
Abstract
Objective Unique long region 83 (UL83) gene is the symbol of human cytomegalovirus (HCMV) reactivated infection. This study is to investigate relationships between UL83 and vulnerable plaques rupture related factors, including angiopoietin (Ang) and vascular endothelial growth factor (VEGF) in HCMV infected human brain vascular smooth muscle cells (HBVSMCs). Methods (1) Cytoflowmetry was used to identify the efficiency of fluorescein amidite (FAM) labeled small interfering RNA (siRNA) transfection into HBVSMCs. (2) Three groups of siRNA were transfected into HBVSMCs infected with HCMV AD169 (MOI=10) in advance, and combinedwith groups of blank, infected and infected but ineffective interfered to find which siRNA could silence UL83 gene to the greatest extent. (3) The messenger ribonucleic acid (mRNA) and protein expressions of Ang-1, Ang-2 and VEGF-A were detected at different time from 0, 6, 12, 24, 48 until 72 hours after HBVSMCs infected with HCMV AD169 (MOI=10), respectively. (4) mRNA and protein expressions of Ang-1, Ang-2 and VEGF-A in blank, infected & effective interfered, infected but ineffective interfered and only infected HBVSMCs were compared. Results A total of 95.95％ of HBVSMCs had been transfected with FAM labelled siRNA after 6 hours. Forty-eight hours after transfected with siRNA, relative expression of UL83 mRNA within HBVSMCs infected with HCMV AD169 (MOI=10) reduced 78.7％ at most, while 72 hours after transfected with siRNA, relative expression of UL83 encoded pp65 protein reduced 81.3％ at most. Relative mRNA and protein expressions of Ang-10, 6, 12, 24, 48 and 72 hours after HBVSMCs infected with HCMV AD169 (MOI=10) gradually reduced (P<0.01), while relative mRNA and protein levels of Ang-2 and VEGF-A gradually increased (P<0.01). After silencing UL83 gene with effective siRNA, both reduction of Ang-1 expression and increase of Ang-2, VEGF-A expressions had been halted. Conclusion HCMV AD169 can reduce Ang-1 expression but increase Ang-2, VEGF-A expressions at the same time. Silence HCMV AD169 UL83 gene expression may halt such trend, which suggests the potential capability of HCMV making target cells producing angiogenic microenvironment through UL83 gene.关键词
人巨细胞病毒UL83基因/小干扰核糖核酸/促血管生成素-1/促血管生成素-2/血管内皮生长因子-AKey words
Human cytomegalovirus UL83 gene/Small interfering RNA/Angiopoietin-1/Angiopoietin-2/Vascular endothelial growth factor-A
