siRNA干扰p38α上调人血管eNOS基因启动子转录活性  

Up-regulation of the transcriptional activity of human eNOS promoter by siRNA interfering with p38α

作　　者：王宝玉[1] 邢飞跃[1] 刘娜[1] 李卓[1] 唐征乐[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实验中心,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2010年第6期539-543,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(30471635;30971465);广东省自然科学基金项目(04010451;5006033);暨南大学"211工程"基金项目(2009)

摘　　要：目的:通过靶向p38α的siRNA干扰人血管内皮细胞的p38α信号通路,探索其对血管内皮型一氧化氮合酶(eNOS)基因启动子转录活性的调节作用。方法:利用W estern印迹检测siRNA-p38α干扰HUVEC-12细胞后p38α蛋白的表达,确定siRNA-p38α的最佳作用时间和浓度,然后通过双荧光素酶报告基因技术检测转染siRNA-p38α后细胞裂解液的荧光强度,分析eNOS基因启动子的转录活性变化。结果:与对照组比较,siRNA-p38α干扰后p38α蛋白表达明显减弱,而相应的eNOS启动子转录活性上调,以100 nmol/L的siRNA-p38α作用48 h的效果最佳。结论:siRNA干扰p38α信号可上调人血管内皮细胞eNOS基因的转录活性。Aim：To investigate the roles of p38α signaling pathway in regulation of the transcription activity of a human endothelial nitric oxide synthase（eNOS） promoter by using p38α-targeting siRNA in a human vascular endothelial cell line（HUVEC-12）. Methods： Expression of p38α protein in the HUVEC-12 cells treated with the siRNA was measured by Western blotting,and the optimal effecting time and optimal concentration of the siRNA were determined.Then,dual-luciferase reporter gene assay was employed to determine the fluorescence intensity of the lysate from the HUVEC-12 cells treated with the siRNA and thereby the activity of the eNOS promoter was analyzed.Results： Compared with the control,the expression of p38α was significantly weakened by interference of p38α-targeting siRNA,and accordingly the corresponding activity of eNOS promoter was up-regulated especially in the group with 100 mM siRNA-p38α for 48 hours.Conclusion：The siRNA interfering with p38α signal can up-regulate the transcriptional activity of the human eNOS promoter.

关 键 词：一氧化氮合酶 p38α丝裂原活化蛋白激酶 小分子干扰RNΑ 内皮细胞 

分 类 号：R392.12[医药卫生—免疫学]