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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。
To investigate the effects of store-operated calcium channels (SOCs) and receptor-activated calcium channels (ROCs) on the calcium sensitivity of human umbilical vein endothelial cells (HUVECs) Receptor (Ca2 +) - sensing receptor (CaR) -mediated Ca2 + influx and NO production were investigated in this experiment by blockade of SOC blockers and nonselective cation channels ROC agonist and ROC blocker were used to detect the changes of intracellular Ca 2+ concentration and [Ca 2+] _i with Fura-2 / AM. NO fluorescence probe DAF -FM DA was used to determine the changes of endothelial nitric oxide synthase (eNOS) activity and NO production. The results showed that compared with the control group (Spermine + Ca 2+ group, CaR), the activities of [Ca 2+] i, eNOS and NO in HUVEC were decreased (P <0.05). However, ROC agonist partially abolished the blockade of ROC blockade (P <0.05). Combined inhibition of SOC and ROC further enhanced the inhibitory effect (P <0.05). The above results indicate that SOC and ROC are involved in CaR-mediated calcium influx and NO production during Spermine stimulation of CaR-mediated [Ca 2+] i, eNOS activity and NO generation.