人冠状动脉内皮细胞活检方法的建立  

Identification of human coronary artery endothelial cells obtained by coronary endovascular biopsy

作　　者：于世勇[1] 黄岚[1] 宋耀明[1] 李爱民[1] 覃军[1] 于学军[1] 耿召华[1] 赵刚[1] 林春梅[1] 陶春蓉[1] 

机构地区：[1]第三军医大学新桥医院全军心血管内科研究所,重庆400037

出　　处：《中华心血管病杂志》2008年第3期240-242,共3页Chinese Journal of Cardiology

基　　金：国家自然科学基金资助项目（30470729）

摘　　要：目的建立人冠状动脉内皮细胞活检方法，为研究内皮细胞在生理及病理状态下的结构和功能提供有益的帮助。方法对28例行冠状动脉介入治疗的患者，应用冠状动脉导丝获取冠状动脉病变部位内皮细胞，利用免疫磁珠法进行分选，瑞氏染色后计数，并通过Ⅷ因子相关抗原抗体免疫荧光法鉴定其内皮源性，通过荧光素乙酰乙酸盐（FDA）和碘化丙啶（PI）双荧光染色法检测细胞膜的完整性，通过对DiI-acLDL的摄取观察细胞的代谢活性。结果平均每位患者可通过导丝获取约96个细胞，活检所得细胞具有内皮细胞的形态学特点，经鉴定是胞膜完整、有活性的内皮细胞。结论通过常规的冠状动脉介入操作，结合免疫磁珠分选，可以在病变血管获得一定数量的有活性的冠状动脉内皮细胞，所得细胞可用于进一步的免疫细胞化学及分子生物学检测，能够准确地反映冠状动脉内皮细胞在不同疾病状态下的功能变化，有助于深入探讨冠状动脉疾病的病理过程。Objective To develope a method to obtain and identify human coronary artery endothelial cells obtained during percutaneous coronary interventions （PCI）. Methods Coronary guide wires were used to obtain endothelial cells from coronary arteries in 28 patients undergoing PCI. The cells were eluted from the wire tips and then purified by magnetic beads coated with anti-CD146 antibody, yon Willebrand factor （vWF） was used as an immunocytochemical marker for endothelial cells. The cellular viability was evaluated by observing cell membrane integrity and energy-dependent uptake of DiI-labeled acetylated low-density lipoprotein. Results An average of 96 coronary artery endothelial cells with good viability per patient were obtained by one guide wire. vWF identification showed their endothelial morphology and immunoreactivity. Conclusion The viable coronary endothelial cells could be obtained during routine percutaneous coronary interventions combined with magnetic beads isolation technique. These cells may be used for further cellular functional analyses （ such as immunocytochemistry and molecular biology ） and expand our understanding on mechanisms of coronary artery diseases.

关 键 词：冠状动脉疾病 内皮 血管 活组织检查 

分 类 号：R686[医药卫生—骨科学]