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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72h[(0.71±0.06)μmol/106 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、72h,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。
Objective To investigate the effect and mechanism of human cytomegalovirus infection on cholesterol metabolism in umbilical artery smooth muscle cells. Methods Smooth muscle cells were isolated from umbilical arteries and challenged with 1 human cytomegalovirus at a multiplicity of infection (MOI) while simulating infection with 3% fetal bovine serum in DMEM / F12 maintenance fluid for use as a control. The change of cholesterol content in smooth muscle cells after virus challenge was detected. The changes of gene expression profile after virus challenge were detected by gene expression microarray. The changes of enzymes related to cholesterol metabolism were investigated and confirmed by fluorescence quantitative RT-PCR. Results The level of intracellular cholesterol was significantly higher than that of the mimic group (P = 0.000 2) at 72 hours after infection ([0.71 ± 0.06] μmol / 106 cells]. At 48 and 72 h after VSMCs were challenged with human cytomegalovirus, the expressions of HMG-CoA synthase and HMG-CoA reductase were significantly upregulated (P <0.01). Conclusion Human cytomegalovirus (CMV) infection may up-regulate the expression of HMG-CoA reductase gene and HMG-CoA reductase gene, resulting in increased cholesterol synthesis in vascular smooth muscle cells and imbalance of intracellular cholesterol metabolism.