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人类巨细胞病毒感染对脐动脉血管平滑肌细胞胆固醇代谢的影响
李玲芳1,陈华2,邬国军1*
0
(. 中南大学湘雅医学院,长沙 410078; 2. 长沙市第三医院检验科,长沙 410015)
摘要:
\[摘要\] 目的 探讨人类巨细胞病毒感染对脐动脉血管平滑肌细胞内胆固醇代谢的影响及机制。方法 自脐动脉分离平滑肌细胞,以1个感染复数(multiplicity of infection,MOI)的人类巨细胞病毒攻击,同时用含3%小牛血清的DMEM/F12维持液进行模拟感染以用作对照。测定病毒攻击后平滑肌细胞内胆固醇含量的变化,以基因表达谱芯片技术检测病毒攻击后细胞基因表达谱的变化,研究与胆固醇代谢有关酶的变化并经荧光定量RTPCR验证。结果 细胞内胆固醇含量在病毒感染后72 h\[(0.71±0.06) μmol/106 cells\]显著高于模拟感染组(P=0.000 2)。脐动脉平滑肌细胞被人类巨细胞病毒攻击后48、72 h,细胞中与胆固醇合成代谢有关的HMGCoA合成酶及HMGCoA还原酶的表达明显上调(P<0.01)。结论 人类巨细胞病毒感染可能通过上调HMGCoA合成酶及HMGCoA还原酶基因的表达水平,导致血管平滑肌细胞内胆固醇合成增加,引起细胞内胆固醇代谢失衡。
关键词:  巨细胞病毒  HMG辅酶A合成酶  HMG辅酶A还原酶  胆固醇  脐动脉  平滑肌细胞
DOI:10.3724/SP.J.1008.2011.0150
投稿时间:2010-10-18修订日期:2011-01-20
基金项目:湖南省卫生厅科研基金(B2008 079).
Influence of human cytomegalovirus infection on cholesterol metabolism in smooth muscle cells of umbilical arteries and the related mechanisms
LI Ling-fang1,CHEN Hua2,WU Guo-jun1*
(1. Xiangya School of Medicine,Central South University,Changsha 410078,Hunan,China; 2. Department of Clinical Laboratory, The Third Hospital of Changsha,Changsha 410015,Hunan,China)
Abstract:
\[Abstract\] Objective To investigate the effect of human cytomegalovirus(HCMV) infection on cholesterol metabolism in smooth muscle cells of the umbilical arteries and the related mechanismsMethods Vascular smooth muscle cells were isolated from umbilical arteries and were challenged with 1 MOI HCMV, and the intercellular content of cholesterol was determined.Cells treated with DMEM/F12 solution containing 3% fetal bovine serum were taken as mock infection controls.The differentially expressed genes were screened by cDNA microarray in each group,and the abnormally expressed genes associated with cholesterol metabolism were identified by fluorescence quantitative RTPCR.Results The intracellular content of cholesterol (\[0.71±0.06\] μmol/106 cells) was significantly higher in HCMV infection group at 72 h after infection compared with that in the control group (t=7.950,P=0.000 2).cDNA microarray and fluorescence quantitative RTPCR showed that the expressions of HMGCoA synthetase and HMGCoA reductase were greatly upregulated compared with those in the mock infection group 48 and 72 h after infection (P<0.01).Conclusion Human cytomegalovirus infection can increase the synthesis of cholesterol through upregulating cholesterol metabolismassociated genes in vascular smooth muscle cells,leading to unbalancd cholesterol metabolism.
Key words:  cytomegalovirus  HMG-CoA synthetase  HMG-CoA reductases  cholesterol  umbilical arteries  smooth muscle cells