动脉粥样硬化大鼠动脉组织中组织蛋白酶D的表达  

Expression of Cathepsin D in Aorta Tissue from Rats with Atherosclerosis

作　　者：张志钢[1] 张晓兰[2] 舒青[3] 郑强荪[1] 

机构地区：[1]第四军医大学唐都医院心血管内科 [2]南京军区福州总医院消化内科,福建省福州市350025 [3]第四军医大学基础部遗传与发育教研室,陕西省西安市710032

出　　处：《中国动脉硬化杂志》2008年第6期461-464,共4页Chinese Journal of Arteriosclerosis

摘　　要：目的用基因芯片技术研究动脉粥样硬化大鼠动脉组织中差异表达的基因,以分析组织蛋白酶D基因表达变化。方法以高脂饲料喂养加维生素D3腹腔注射建立大鼠动脉粥样硬化模型,测定血清总胆固醇及甘油三酯;病理学观察HE及油红O染色的冠状动脉及主动脉;提取主动脉RNA作基因芯片检查,用免疫荧光及免疫印迹法检测组织蛋白酶D在动脉中表达的分布情况及表达水平。结果14周时模型组总胆固醇和甘油三酯较对照组明显升高(P均<0.01);病理学观察发现模型组形成纤维增生性动脉粥样硬化病变;基因芯片结果显示与对照组比较,模型组组织蛋白酶基因表达上调,组织蛋白酶D表达上调,R值(Cy3/Cy5)为2.14(P<0.01);免疫荧光检测发现模型组中组织蛋白酶D表达主要分布在动脉内膜、中膜;免疫印迹法检测动脉中组织蛋白酶D蛋白显示模型组表达较对照组增加(P<0.05)。结论组织蛋白酶及其内源性抑制剂的失衡可能是导致动脉粥样硬化的因素之一,组织蛋白酶D可能参与了血管重塑及细胞凋亡从而促进动脉粥样硬化的发展。Aim To investigate the potential candidate genes related to extracellular matrix （ECM） which might play roles in atherosclerosis by microarray technology. Methods Sixteen 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, 0.5% sodium cholate, 5% refined sugar, 10% lard, and 81.5% base feed） （model group, n = 8）. After 14 w, serum total cholesterol （TC） and triglyeride （TG） 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 expression was measured by immunofluorescence and western blot. Results Compared with the control group, model group showed elevated serum TC and TG （ both P 〈 0.05 ） and aortic fibrous plaque formation. 510 genes were up-regulated and 1 218 ones were down-regulated in aorta tissue of model group by DNA microarray analysis when compared with coutrol group. Further immunohistochemistry and western blot analysis revealed cathepsin D protein expression was significantly increased in aorta of model group whereas normal group showed line or no immuno-detectable cathepsin D expression. Conclusion Cathepsin D might influence vascular remodeling, which plays an important role in As.

关 键 词：内科学 动脉粥样硬化模型 大鼠 组织蛋白酶D 基因芯片 免疫荧光 免疫印迹 

分 类 号：R5[医药卫生—内科学]