为探讨人脐静脉内皮细胞(HUVEC)游离钙离子浓度是否受细胞外钙受体(CaR)的调节。采用Westernblot及RT-PCR技术检测CaR在HUVEC中蛋白及mRNA表达。使用CaR激动剂精胺,负性变构调节剂Calhex231,通过钙荧光探针(Fura-2/AM)负载HUVEC检测细胞内钙离子浓度([Ca2+]i)。结果显示:(1)HUVEC中有CaR的表达。(2)在Fura-2/AM负载HUVEC中加入精胺刺激,[Ca2+]i为△ratio=0.24±0.01,先加入Calhex231作用1min后再加入Calhex231+精胺刺激,[Ca2+]i为△ratio=0,两组比较差异有统计学意义(P〈0.05)。由此可知,CaR调节HUVEC游离钙离子浓度。
To study whether intracellular Ca2+concentration ([Ca2+]i) is regulated by Ca-sensing receptor (CaR) in human umbilical vein endothelial cells(HUVEC).CaR protein was demonstrated by Western blot.The mRNA expression of CaR was investigated by RT-PCR in HUVEC.The fluorescence Ca2+ indicator Fura-2 /AM was used to observe [Ca2+]i in the presence of CaR agonists spermine and negative allosteric modulator Calhex231.(1) CaR is present in HUVEC.(2)There was significantly difference in [Ca2+]i between spermine (ratio=0.24 ±0.01) and Calhex231+spermine (ratio=0) (P0.05).CaR regulates [Ca2+]i in HUVEC.