摘要
为了探讨钙库活化的钙通道(store-operated calcium channels,SOC)和受体活化的钙通道(receptor-operated calcium channels,ROC)在人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)钙敏感受体(Ca^(2+)-sensing receptor,CaR)介导的钙内流及NO生成中的作用和机制,本实验通过分别或联合使用SOC阻断剂、非选择性的阳离子通道阻断剂、ROC激动剂和ROC阻断剂,采用Fura-2/AM检测细胞内钙离子浓度(intracellular Ca^(2+)concentration,[Ca^(2+)]_i)变化,NO荧光探针DAF-FM DA测定细胞内内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)活性和NO生成的变化。结果显示,与对照组(Spermine+Ca^(2+)组,激活CaR)相比,单独阻断SOC或ROC,HUVEC中[Ca^(2+)]_i、eNOS活性和NO含量均降低(P<0.05),而ROC激动剂可部分取消ROC阻断剂的阻断作用(P<0.05),SOC和ROC联合阻断可进一步加强上述抑制作用(P<0.05)。上述结果表明,在Spermine刺激激活CaR介导[Ca^(2+)]_i、eNOS活性和NO的生成过程中,SOC和ROC以协同的方式参与了CaR介导的钙内流及NO生成。
This paper aims to investigate the effect of store-operated calcium channels （SOC） and receptor-operated calcium channels （ROC） on Ca2＋-sensing receptor （CaR）-induced extracellular Ca2＋ influx and nitric oxide （NO） generation in human umbilical vein endothelial cells （HUVEC）. SOC blocker, non-selective cation channel blocker, ROC agonist and ROC blocker were used separately and combined. Intracellular Ca2~ concentration （[Ca：~]i） was measured by Fura-2/AM loading. The activity of endothelial nitric oxide synthase （eNOS） and the production of NO were determined by the DAF-FM diacetate （DAF-FM DA）. The results showed that increases of [Ca2＋]~, eNOS activity and NO generation induced by CaR agonist Spermine were all reduced after single blocking the SOC or ROC, respectively （P 〈 0.05）. ROC agonist can partially abolish the ROC blocker＇s effect （P 〈 0.05）. The above mentioned effects evoked by CaR agonist Spermine were further reduced when blocking both SOC and ROC than single blocking SOC or ROC in HUVEC （P 〈 0.05）. In conclusion, these results suggest that the SOC and ROC participate in the processes of CaR-evoked extra-cellular Ca2＋ influx and NO generation by a synergistic manner in HUVEC.
出处
《生理学报》
CAS
CSCD
北大核心
2013年第5期553-561,共9页
Acta Physiologica Sinica
基金
supported by the National Natural Science Foundation of China(No.31160239)
关键词
内皮细胞
钙敏感受体
钙信号
NO
钙库活化的钙通道
受体活化的钙通道
endothelial cells
calcium-sensing receptor
calcium signaling
nitric oxide
store-operated calcium channels
receptor-operated calcium channels
