【摘要】 前期研究发现小凹蛋白-1(caveolin-1,Cav-1)和细胞外钙敏感受体(extracellular Ca2+-sensing receptor,CaR)在人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)膜上有共定位,且Cav-1下调CaR介导的钙内流。本实验探讨HUVECs中Cav-1与CaR的亚细胞定位,可为进一步机制研究提供理论依据。以3~4代HU-VECs随机分为2组:(1)对照组;(2)甲基-β-环糊精(MβCD)处理组,分别采用蔗糖密度梯度离心和Western blot技术提取含小凹膜碎片和检测Cav-1和CaR蛋白的表达。结果显示,(1)MβCD处理组HUVECs细胞中Cav-1和CaR蛋白的表达与对照组相比无差异(P>0.05)。(2)MβCD处理组小凹区Cav-1和CaR蛋白的表达低于对照组(P<0.05)。由此可知,Cav-1和CaR蛋白共定位于HUVECs的小凹区,MβCD可抑制两者在小凹的共定位。更多还原

【Abstract】 Our previous results demonstrated that extracellular Ca2+-sensing receptor(CaR) and caveolin-1(Cav-1) co-localized on the plasma localization of Cav-1 and CaR in HUVECs,which provide a theoretical basis for further functional studies.Methods:The third or fourth passage of HUVECs were randomly divided into two groups:(1) Control group;(2)methyl-β-cyelodextrin(MβCD) group.We isolated caveolae-enriched membrane fractions from MβCD-treated HUVECs by TritonX-100 sucrose density gradient fractionation and detected the expressions of Cav-1 and CaR protein by Western blot,respectively.Results:(1)The expression of Cav-1 and CaR proteins between control group and MβCD group was no difference(P>0.05).(2)The expressions of Cav-1 and CaR proteins in caveolae after MβCD treated was significantly reduced(P<0.05).Conclusion:The present study suggests that CaR and Cav-1 co-localize in caveolae in HUVECS,MβCD reduces the co-localization of Cav-1 and CaR in caveolae.更多还原