Study of atherosclerosis using DNA microchip

采用聚合酶链反应、Southern印迹和原位杂交技术对33例主动脉组织的单纯疱疹病毒1型（HSV-1）、eb病毒（EBV）和巨细胞病毒（CNV）的核酸进行了检测。23例死亡时年龄从23周龄到75岁不等。组织学上组织无动脉粥样硬化。10例死亡时年龄从53岁到75岁不等，为动脉粥样硬化性主动脉组织。在非动脉粥样硬化组中没有出现双重或三重感染，而在动脉粥样硬化组中，10人中有6人对两种病毒呈阳性，10人中有2人对三种病毒呈阳性。病毒定位于与内皮细胞和平滑肌细胞形态一致的细胞中。接下来我们调查了肺炎衣原体和沙眼衣原体感染。40%的组织检查呈肺炎原体阳性，而非动脉粥样硬化性主动脉则没有。电镜下在动脉粥样硬化主动脉内膜巨噬细胞样细胞中可见肺炎原体。在动脉粥样硬化和非动脉粥样硬化主动脉均未发现沙眼衣原体。在此基础上，我们进一步研究了体外培养的内皮细胞、平滑肌细胞和巨噬细胞感染疱疹病毒和肺炎C后基因表达的变化。内皮细胞最易受感染，并在疱疹感染后6小时开始退化。虽然形成了多核巨细胞，但经过5天的感染后，所有细胞最终都出现了溶解。肺炎支原体感染表现为持续感染。约有30个基因高表达，其中约有10个基因与胆固醇代谢有关。热休克蛋白是与感染因子和动脉粥样硬化有关的最重要的关键基因。

The viral nucleic acid of herpes simplex virus type 1 (HSV-1), Epstein-Barn virus (EBV) and Cytomegalovirus (CNV) was studied by polymerase chain reaction, Southern blotting and in situ hybridization in aortic tissues from 33 autopsies. 23 cases ranged from 23 weeks to 75 years of age at the time of death. And the tissue was histologically non-atherosclerotic. 10 cases ranged from 53 years to 75 years old at death were atherosclerotic aortic tissue. Neither double nor triple infections occurred in the non-atherosclerotic group, whereas six of 10 were positive for two viruses, and two of 10 were positive for three viruses in the atherosclerotic group. The viruses were localized in cells morphologically consistent with endothelial cells and smooth muscle cells. Next we investigated both Chlamydia pneumoniae and Chlamydia trachomatis infections. 40% of the tissue examined were positive for C. pneumoniae in contrast to absence in non-atherosclerotic aorta. Elementary bodies of C. pneumoniae were found in macrophage-like cells in the intima of atherosclerotic aorta by electron microscopy. C. trachomatis was not found in both atherosclerotic and non-atherosclerotic aorta.Based on these results, we further studied the changes of gene expression in cultured endothelial cells, smooth muscle cells and macrophages when they were infected with Herpes virus and C, pneumoniae. Endothelial cells were most susceptible and begin to degenerate lytic 6 hours after the herpes infection. Multinucleated giant cells were formed, but all the cells finally showed lytic after 5 clays of infection. C. pneumoniae infection, however, revealed persistent infection. About 30 genes were expressed high and about 10 genes of them were related to cholesterol metabolism. Heat shock proteins was the most important key gene in relation to infectious agents and antherosclerosis.