大鼠冠状动脉粥样硬化钙化组织与c-kit^(+)和sca-1^(+)细胞的关系研究  

Relationship between calcified tissue of coronary atherosclerosis and c-kit^(+)and sca-1+cells in rats

作　　者：孙攀文 郭永龙 贾阳阳[1] 孙昌业 郭志坤[1] Sun Panwen;Guo Yonglong;Jia Yangyang;Sun Changye;Guo Zhikun(Henan Key Laboratory of Medical Tissue Regeneration,Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Department of Cardiology,the First Affiliated Hospital of Xinxiang Medical Hospital,Xinxiang 453003,Henan Province,China)

机构地区：[1]新乡医学院河南省医用组织再生重点实验室,河南新乡453003 [2]新乡医学院第一附属医院心内科,河南新乡453003

出　　处：《中国临床解剖学杂志》2023年第4期453-458,共6页Chinese Journal of Clinical Anatomy

基　　金：国家自然科学基金(81570268);河南省科技攻关(222102310094)。

摘　　要：目的探讨大鼠冠状动脉粥样硬化钙化区域sca-1、c-kit的表达变化,为进一步研究冠状动脉粥样硬化机制奠定细胞学基础。方法30只健康成年SD大鼠,随机分为正常对照组(n=8)和动脉粥样硬化钙化模型组(n=22)。采用丙基硫氧嘧啶灌胃、高脂高胆固醇饲料喂养和维生素D3腹腔注射方法制备模型。模型制备成功后,通过免疫组织化学和免疫荧光技术观察sca-1、c-kit在模型组与正常组的大鼠冠状动脉组织中的表达变化。利用免疫荧光双标技术证明sca-1与α-SMA的共定位表达。利用western blotting技术检测模型组与正常组的冠状动脉组织c-kit、sca-1蛋白的表达差异。结果造模12周,大鼠冠状动脉管壁明显增厚,内膜明显增生,平滑肌纤维排列显著紊乱,坏死物质沉积、脂质沉积和钙结节沉淀。免疫组化和免疫荧光结果显示sca-1+细胞和c-kit^(+)细胞在动脉粥样硬化钙化区域的冠状动脉内膜、中膜、外膜中数量明显高于正常对照。病变区域的部分细胞存在sca-1与α-SMA的共表达。Western blotting结果显示动脉粥样硬化和钙化区域的c-kit、sca-1蛋白的表达量高于正常对照组。结论sca-1+细胞和c-kit^(+)细胞有向冠状动脉粥样硬化钙化区域迁移的倾向,其机制尚需研究。Objective To investigate the expression change of sca-1 and c-kit in the calcified area of coronary atherosclerosis in rats,so as to provide clinical reference for further study of the mechanism of coronary atherosclerosis.Methods Thirty-two healthy adult SD rats were randomly divided into a normal control group(n=8)and an atherosclerotic calcification model group(n=24).The model was prepared by gavage of propylthiouracil,feeding with high fat and cholesterol diet and intraperitoneal injection of vitamin D3.After the model was successfully prepared,the expression changes of sca-1 and c-kit in the coronary artery tissues of the model group and the normal group were observed by immunohistochemistry and immunofluorescence techniques.Immunofluorescence technique was used to detect the co-expression of sca-1 andα-SMA.Western blotting was used to detect the differences of c-kit and sca-1 protein expression between model group and normal group.Results At the 12th week of modeling,the coronary artery wall of rats was significantly thickened,the intima was significant hyperplasia,the smooth muscle cells were arranged in a significant disorder,and there were necrotic substance deposition,lipid deposition and calcium nodule precipitation.Immunohistochemical and immunofluorescence results showed that sca-1+cells and c-kit^(+)cells were significantly higher in the intima,media and adventitia of coronary artery in atherosclerotic and calcified areas than those in the normal control group.Some cells have co-expression of sca-1 andα-SMA in the location of lesion.Western blotting result showed that the expressions of c-kit and sca-1 protein in atherosclerotic and calcified areas were higher than those in normal control group.Conclusions Sca-1+cells and c-kit^(+)cells have the tendency to migrate to the calcification area of coronary atherosclerosis,and the mechanism needs to be further investigated.

关 键 词：SCA-1 C-KIT 动脉粥样硬化钙化 免疫组织化学 大鼠心脏 

分 类 号：R361.2[医药卫生—病理学]