氯化钴对人eNOS基因启动子转录活性的影响  

Effect of cobaltous chloride on the transcription activity of human eNOS promoter

作　　者：樊振华[1] 邢飞跃[1] 

机构地区：[1]暨南大学生命科学技术学院组织移植与免疫中心,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2009年第4期379-382,共4页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金资助项目(30471635);广东省自然科学基金资助项目(04010451;5006033)

摘　　要：目的:建立氯化钴诱导的人脐静脉血管内皮细胞-12(HUVEC-12)缺氧模型,研究人血管内皮细胞一氧化氮合酶(eNOS)基因的启动子活性变化。方法:用含不同浓度氯化钴的培养基培养细胞,检测细胞培养上清中的乳酸脱氢酶(LDH)含量;将已构建的pGL2-eNOS-p质粒转染HUVEC-12细胞,利用双荧光素酶报告基因技术检测在不同浓度氯化钴和不同作用时间下的eNOS启动子转录活性。结果:氯化钴刺激下HUVEC-12细胞培养上清的LDH含量随氯化钴作用浓度增加而提高;氯化钴刺激后转染细胞的eNOS启动子活性呈现随氯化钴剂量增加而升高的趋势,随作用时间的延长而上升的趋势。结论:成功建立氯化钴诱导HUVEC-12细胞缺氧的体外模型。Aim:To establish a hypoxia model of human umbilical vein endothelial cells-12(HUVEC-12) with stimulating of Cobaltous chloride in vitro,and investigate activity of human endothelial nitric-oxide synthase(eNOS) promoter. Methods: The cells were treated with the different concentrations of Cobaltous chloride.The lactic acid dehydrogenase(LDH) from the supernatant of the cells was detected by a biochemical analysis.The cells were co-transfected with a pGL2-eNOS-p vector while stimulated with Cobaltous chloride at the different concentrations and disposing time,and the transcription activity of human eNOS promoter was determined through using a double luciferase reporter gene system.Results: As the concentration of cobaltous chloride increased,the LDH and the promoter transcription activity were also increased.Furthermore,there was a tendency that the transcription activity increased as time lasted.Conclusion:These results suggest that in vitro hypoxia model of HUVEC-12 cells induced by Cobaltous chloride has been successfully established.

关 键 词：一氧化氮合酶 启动子 乳酸脱氢酶 氯化钴 内皮细胞 

分 类 号：R392.12[医药卫生—免疫学]