The temporal correlation of IL-1b、IL-1ra、ICAM-1 and VCAM-1 expressions during the development of atherosclerosis

• Main Authors: Yu-Min Chien, 簡玉敏
• Other Authors: Yuh-Lien Chen
• Format: Others
• Language: zh-TW
• Published: 2000
• Online Access: http://ndltd.ncl.edu.tw/handle/70284359152549790926

碩士 === 國立陽明大學 === 解剖學研究所 === 88 === ABSTRACT Neointimal hyperplasia, mainly the proliferation of smooth muscle cells and accumulation of macrophage-derived foam cells and interleukin-1 beta might play a central role in the process of atherosclerosis. In animal study, we investigated the expression of IL-1beta, IL-1ra, ICAM-1 and VCAM-1 during atherosclerosis in high cholesterol-fed rabbits by in situ hybridization, immunohistochemical staining and western blot. New Zealand White rabbits were fed with 2% cholesterol-containing diet for 2 week, 3 weeks and 6 weeks. At designated time points, artery were excised for paraffin embedding. In situ hybridization using digoxigenin-labeled IL-1beta and IL-1ra cDNA as a probe was conducted to determine the distribution of IL-1beat and IL-1ra mRNA. Immunohistochemical staining was used to study the expression of IL-1beta, IL-1ra, ICAM-1 and VCAM-1 and to identify cells in the arterial wall responsible for IL-1beta, IL-1ra, ICAM-1 and VCAM-1 expression. Western blot was used to determine the volume of IL-1beta and IL-1ra. Otherwise, we used the methods of above to determine the expression of IL-1beta, IL-1ra, ICAM-1 and VCAM-1 in human serious atherosclerosis. In cell cultured study, we observed the expression of ICAM-1 and VCAM-1 in IL-1beta-treated HASMCs by cell ELISA. The results revealed : IL-1beta mRNA expression was detected in the thickened intima in cholesterol-fed rabbit for 2 weeks, 3 weeks and 6 weeks and in human atherosclerosis. The thickened intima also showed strong IL-1beta protein reactions. IL-1ra mRNA expression was detected in the slight thickened intima in cholesterol-fed rabbit for 2 weeks and 3 weeks and IL-1ra mRNA expression was remarkable in the thickened intima in cholesterol-fed rabbit for 6 weeks and in human atherosclerosis. But the expression of IL-1ra protein was not clear and obvious. The expression of ICAM-1 and VCAM-1 protein were remarkable in the thickened intima in cholesterol-fed rabbit for 6 weeks and in human atherosclerosis. By using the antibodies against the macrophages and smooth muscle cells respectively,the cells in the thickened intima were considered to be macrophages and few smooth muscle cells in cholesterol-fed rabbits. Nevertheless, the major component of the thickened intima is smooth muscle cell in human serious atherosclerosis. The expression of ICAM-1 and VCAM-1 protein were not increase in 10ng/ml IL-1beta-treated HASMCs in this study.