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钙库活化的钙通道和受体活化的钙通道参与人脐静脉内皮细胞钙敏感受体介导的钙内流和NO生成

赵慧, 梁霄, 钟华, 张春军, 何芳

新疆石河子大学医学院病理生理教研室/新疆地方与民族高发病教育部重点实验室，石河子 832002；新疆医科大学附属肿瘤医院淋巴瘤与乳腺内科，乌鲁木齐 830011

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(Ca2+-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 Ca2+ concentration, [Ca2+]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 + Ca2+组，激活CaR)相比，单独阻断SOC或ROC，HUVEC中[Ca2+]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[Ca2+]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。

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

分类号：R363

[Involvement of store-operated calcium channels and receptor-operated calcium channels in Ca2+-sensing receptor-evoked extracellular Ca2+ influx and NO generation in human umbilical vein endothelial cells.] [Article in Chinese]

ZHAO Hui, LIANG Xiao, ZHONG Hua, ZHANG Chun-Jun, HE Fang

Department of Pathophysiology/Key Laboratory of Education Ministry of Xinjiang Endemic and Ethnic Diseases, Medical College of Shihezi University, Shihezi 832002, China； Department of lymphoma and breast medicine, Affiliated Tumor Hospital, Xinjiang Medcial University, Urumqi 830011, China

Abstract

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 ([Ca²⁺]_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⁺]_i, 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 extracellular Ca²⁺ influx and NO generation by a synergistic manner in HUVEC.

Key words: Endothelial cells; calcium-sensing receptor; calcium signaling; nitric oxide; store-operated calcium channels; receptor-operated calcium channels

收稿日期：2013-02-27　　录用日期：2013-08-16

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引用本文：

赵慧, 梁霄, 钟华, 张春军, 何芳. 钙库活化的钙通道和受体活化的钙通道参与人脐静脉内皮细胞钙敏感受体介导的钙内流和NO生成[J]. 生理学报 2013; 65 (5): 553-561.

ZHAO Hui, LIANG Xiao, ZHONG Hua, ZHANG Chun-Jun, HE Fang. [Involvement of store-operated calcium channels and receptor-operated calcium channels in Ca2+-sensing receptor-evoked extracellular Ca2+ influx and NO generation in human umbilical vein endothelial cells.] [Article in Chinese]. Acta Physiol Sin 2013; 65 (5): 553-561 (in Chinese with English abstract).