Study of atherosclerosis using DNA microchip

应用聚合酶链反应、Southern印迹和原位杂交技术，对33例尸检主动脉组织中单纯疱疹病毒Ⅰ型（HSV-1）、EB病毒（EBV）和巨细胞病毒（CNV）的病毒核酸进行了研究。23例病例死亡时年龄为23周至75岁。组织学上没有动脉粥样硬化。10例患者死亡时年龄53 ~ 75岁，均为主动脉粥样硬化。在非动脉粥样硬化组中既没有双重感染也没有三重感染，而在动脉粥样硬化组中，10个中有6个对两种病毒呈阳性，10个中有2个对三种病毒呈阳性。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，我们研究了肺炎衣原体和沙眼衣原体感染。40%的受检组织C.肺炎，而非动脉粥样硬化性主动脉中不存在。C.基本体电镜下可见动脉粥样硬化主动脉内膜巨噬细胞样细胞。C.在此基础上，我们进一步研究了培养的内皮细胞、平滑肌细胞和巨噬细胞感染疱疹病毒和肺炎衣原体后基因表达的变化。内皮细胞最为敏感，在疱疹感染后6小时开始变性溶解。多核巨细胞形成，但感染5天后，所有细胞均出现溶解。C.然而，肺炎感染显示持续感染。约有30个基因高表达，其中约10个基因与胆固醇代谢相关。热激蛋白是与病原菌和花药发育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.