哇巴因抑制钠泵β1亚单位和VE-cadherin减弱血管内皮细胞连接功能  

Ouabain reduces the function of cell junction in vascular endothelial cells through inhibiting the expression of Na^+,K^+-ATPase β1-subunit and VE-cadherin

作　　者：徐瑞成[1] 王娜[1] 徐忠伟[1] 陈雪芬[1] 呼文亮[1] 

机构地区：[1]中国人民武装警察部队医学院.天津市职业和环境危害生物标志物重点实验室,天津300162

出　　处：《中国药理学通报》2008年第8期1093-1098,共6页Chinese Pharmacological Bulletin

基　　金：天津市自然科学基金资助项目(No06YFJMJC10400)

摘　　要：目的探讨钠泵抑制剂哇巴因对血管内皮细胞连接的影响及机制。方法以人脐静脉内皮细胞(HUVECs)为靶细胞,Hoechst33342/PI双荧光染色法观察哇巴因作用后细胞凋亡或坏死特征,透射电镜和光镜观察细胞形态结构变化。应用半定量聚合酶链反应检测钠泵α1亚单位、β1亚单位、VE-cadherin和SnailmRNA的表达。结果0.1μmol.L-1哇巴因作用HUVECs24～48h,细胞死亡以凋亡为主,10μmol·L-1哇巴因作用24h,引起细胞坏死;对照组细胞间的细胞连接数量多,结构清晰,而经哇巴因作用后,细胞连接丧失,细胞脱落。哇巴因作用HUVECs后,钠泵α1亚单位和Snail表达上调,β1亚单位和VE-cadherin表达下降,其改变均呈剂量和时间依赖性。结论哇巴因通过下调血管内皮细胞钠泵β1亚单位和VE-cadherin的表达使细胞连接功能减弱。Aim To study the effect of ouabain, Na ＋ , K ＋ -ATPase inhibitor, on cell junction in human um- bilical vascular endothelial cells （ HUVECs ） and involved mechanisms. Methods The feature of cell death was studied by Hoechst33342/PI staining. The morphological changes were observed by light microscopy and transmission electron microscopy. The mRNA expression of Na＋ , K＋-ATPase α1-subunit,β1-sub- unit, VE-cadherin and Snail was examined by reverse transcription polymerase chain reaction （RT-PCR）. Results When treated with 0. 1 μmol ＋ L-＋ ouabain for 24 ＋ 48 hours, the HUVECs cells showed obviously features of apoptosis. Treatment of 10 μmol · L-1 oua- bain for 24 hours could stimulate the necrosis of HU- VECs. The cell junctions were clearly observed in con- trol group of HUVECs, but the cells showed obvious defluxion and loss of cell junctions when treated with 0.1 ＋mol ＋ L-1 ouabain for 48 hours. The expressions of Na ＋ , K ＋ -ATPase otl-subunit and Snail were signifi- cantly up-regulated, and the expressions of β1-subunit and VE-cadherin were significantly down-regulated by ouabain in HUVECs, which showed a dose and time- dependent manner. Conclusion Ouabain could reduce the function of cell junctions in vascular endothe- lial cells by down-regulating the expression of Na ＋, K ＋ -ATPase β1-subunit and VE-cadherin.

关 键 词：哇巴因 血管内皮细胞 细胞连接 钠泵 VE-CADHERIN SNAIL 

分 类 号：R322.123[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]