摘要
目的:用基因芯片技术研究动脉粥样硬化大鼠与正常大鼠动脉组织中差异表达的基因,以探讨动脉粥样硬化的发病机制。方法:16只雄性SD大鼠随机分组,正常对照组8只给予正常饮食及生理盐水腹腔注射,模型组8只大鼠喂食高脂饲料(3%胆固醇、0.5%胆酸钠、5%精制糖、10%猪油、基础饲料81.5%)及维生素D3腹腔注射,实验14周处死动物,测定血浆总胆固醇(total cholesterol,TC)及甘油三酯(triglyeride,TG),取主动脉进行HE染色及油红染色进行病理学观察,提取主动脉RNA作基因芯片检查,以免疫组织化学方法检测组织蛋白酶D(CathepsinD,Cat-D)蛋白表达情况。结果:与正常对照组相比,模型组血浆TG和TC升高(P<0.01),HE及油红染色可见主动脉形成纤维增生性动脉粥样硬化。芯片结果显示,与空白对照组相比,模型组Cat-D基因表达上调。进一步免疫组化检查发现,正常对照组大鼠动脉组织中Cat-D不表达或者低表达,模型组中Cat-D表达明显增加(P<0.01)。结论:AS的发病是多因素作用的多基因改变而引发的复杂病变,组织蛋白酶D可能与其他具有降解基质作用的酶协同参与了血管重塑从而促进AS的发展。
Objective： To investigate the pathogenesis of atherosclerosis （AS） in rats by microarray technology. Methods： 16 male SD rats were randomly subjected to the following treatments： standard diet with saline injection i.p. （control group, n=8 ） or fat diet with vitamin D3 injection i.p. （containing 3% cholesterol, 0.2% sodium cholate, 0.5% refined sugar, 10% lard, and 86.5% base feed） （model group, n=8 ）. After 14 weeks, total cholesterol （TC） and triglyeride （TG） in serum were detected. Thoracic aorta was stained with HE and a solution of oil red O to visualize the lesion area. Total RNA was isolated from the aorta for microarray to explore the differential gene expression profiling. Cathepsin D was measured by immunohistochemistry. Results： Compared with the control group, model group showed elevated TC and TG in serum （both P〈0.05） and aortic fibrous plaques. Aortic cathepsin D of model group was up regulated in gene expression profiling compared with that of the control group. Further immunohistochemistry analysis revealed that cathepsin D protein expression was significantly increased in aorta of model group whereas normal group showed little or no immuno-detectable cathepsin D expression. Conclusions： Cathepsin D mRNA and protein expression were increased in atherosclerostic rats, which indicates that cathepsin D may play an important role in the pathgcathepsin agenesis of AS.
出处
《现代生物医学进展》
CAS
2008年第2期252-254,251,共4页
Progress in Modern Biomedicine
