Study of atherosclerosis using DNA microchip

应用聚合酶链式反应、Southern杂交和原位杂交技术对33例尸检标本的主动脉组织中单纯疱疹病毒1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例死亡时年龄在23周至75岁之间。组织学上无动脉粥样硬化。死亡10例，年龄53~75岁，均为动脉粥样硬化性主动脉组织。非动脉粥样硬化组无二重或三重感染，而动脉粥样硬化组10例中有6例2种病毒阳性，2例3种病毒阳性。病毒定位于细胞内，在形态上与内皮细胞和平滑肌细胞一致。接下来，我们调查了肺炎衣原体和沙眼衣原体感染。接受检查的组织中有40%的组织肺炎衣原体阳性，而非动脉粥样硬化的主动脉组织中没有肺炎衣原体。电子显微镜下可见动脉粥样硬化血管内膜巨噬细胞样细胞内可见肺炎衣原体基本体。在此基础上，我们进一步研究了培养的血管内皮细胞、血管内皮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。感染疱疹病毒后6h，内皮细胞最敏感，开始变性溶解。感染5次后，可形成多核巨细胞，但所有细胞均呈裂解状态。然而，肺炎衣原体感染显示持续感染。约30个基因高表达，其中约10个基因与胆固醇代谢有关。热休克蛋白是与感染性疾病和动脉粥样硬化相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.