TRPC_3参与缺氧诱导人脐静脉血管内皮细胞凋亡  

Effect of TRPC3 on apoptosis of HUVECs induced by hypoxia

作　　者：王嵘[1] 苏晓灵[2] 耿排力[1] 朱德锐[1] 龙启福[1] 孙伟[1] 赵延礼[1] 

机构地区：[1]青海大学医学院生物化学与分子生物学教研室,西宁810001 [2]青海省人民医院心血管科

出　　处：《陕西医学杂志》2009年第12期1598-1603,共6页Shaanxi Medical Journal

基　　金：国家人事部留学回国人员科技活动项目基金资助

摘　　要：目的:探讨瞬时受体电位通道蛋白3(TRPC3)在缺氧诱导人脐静脉血管内皮细胞(HUVECs)凋亡中的作用。方法:采用HUVECs细胞株,复苏传代后计数稀释,接种于6孔培养板中,实验组分为常氧对照组(5%CO2+95%空气)和缺氧组(1%O2+5%CO2+94%N2),处理24h后,应用实时荧光定量RT-PCR法及WesternBlot法检测TRPC3mRNA和蛋白水平的表达变化。构建靶向TRPC3基因的siRNA和无关序列质粒表达载体,分别转染至HUVECs,缺氧处理后再用四甲基偶氮唑盐比色法(Methods the tetrazolium,MTT)检测各组细胞存活率,Hoechest33342荧光染色观察细胞凋亡。结果:缺氧处理24h,HUVECs中TRPC3mRNA和蛋白表达明显升高,细胞存活率为67.4%,与常氧对照组有显著性差异(P<0.05);缺氧+TRPC3干扰组细胞存活率为92.7%,与缺氧组有显著性差异(P<0.05);缺氧+TRPC3假干扰组与缺氧组无显著性差异(P>0.05)。荧光显微镜下观察,常氧对照组胞核均匀蓝染;缺氧处理组和缺氧+TRPC3假干扰组胞核明显固缩、凝聚,可见凋亡小体;而缺氧+TRPC3干扰组胞核均匀蓝染。结论:TRPC3参与缺氧诱导HUVECs凋亡。Objective： To investigate the effect of TRPC3 on apoptosis of human umbilical vein endothelial cells （HUVECs） induced by hypoxia. Methods： Cultured HUVECs were randomly assigned to hypoxia group （1 %O2 ＋ 5 %CO2 ＋ 94 %Nz） and control group （ordinary culture） for 24 h. The expressions of TRPC3 mRNA and protein were detected by real time PCR and Western blot, respectively. The siRNA targeting TRPC3 was constructed and transfected into HUVECs . The viability of the HUVECs was determined by MTT assay, and the morphological changes of the cell nuclei were observed by Hoechst 33342 staining. Results： The expression of TRPC3 was increased significantly after hypoxia treatment for 24 h at mRNA level and protein level. The survival rate of HUVECs （67. 4%） was significantly decreased compared with the untreated cells （P〈0.05）. TRPC3 siRNA treatment significantly increased the survival rate of the hypoxiatreated cells （92.7%, P〈0. 05）, while TRPC3 false-siRNA treatment did not obviously affect the survival of the cells compared with hypoxia treatment （P〈 0. 05）. Under fluorescence microscope, the nuclei of the cultured HUVECs were uniformly blue-stained in ordinary culture and TRPC3 siRNA culture. The cells treated with hypoxia and TRPC3 false siRNA exhibited such apoptotic morphological features as cell shrinkage, condensation, aggregation of nuclear chromatin and presence of bright blue-stained apoptotic body. Conclusion： TRPC3 channel is involved in hypoxia-induced apoptosis of cultured HUVECs.

关 键 词：内皮 血管/病理生理学 蛋白质类/代谢 细胞凋亡 脐静脉 @TRPC3 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R543.5[医药卫生—基础医学]