钙库调控的钙离子通道对大鼠膀胱平滑肌收缩性调控的研究  

Store operated calcium channels regulate contraction of rat bladder smooth muscle in vitro

作　　者：赵滨[1] 白新宇[2] 董兴有 王青青[2] 李龙坤[2] 宋波[1] 

机构地区：[1]第三军医大学西南医院泌尿外科,重庆400038 [2]第三军医大学新桥医院泌尿外科,重庆400037

出　　处：《第三军医大学学报》2014年第11期1178-1182,共5页Journal of Third Military Medical University

摘　　要：目的分析胞内钙库调控的钙离子通道(store operated calcium channels,SOCCs)在大鼠膀胱平滑肌中的功能。方法 RT-PCR检测SOCCs相关蛋白基因表达;使用SOCCs的激动剂CPA(10μmol/L)以及抑制剂SKF-96365(10μmol/L),采用离体肌条的方法观察其肌肉的收缩,全细胞膜片钳技术观察膀胱平滑肌细胞电流,激光共聚焦显微镜观察其膀胱平滑肌细胞内钙离子变化。结果 RT-PCR检测到TRPC3、TRPC4和STIM1基因表达。加入CPA激活SOCCs后,离体肌条实验结果显示膀胱平滑肌离体肌条收缩频率为(5.937 5±1.108 0)/min,比加入CPA前明显增加(P<0.05),收缩幅度为(0.067 1±0.027 0)g,比加入CPA前明显降低(P<0.05);膜片钳实验结果显示钙电流较加入CPA前显著增加,-80 mV电流密度为(-3.127 1±0.908 7)pA/pF,20 mV电流密度为(1.048 1±0.322 5)pA/pF(P<0.05);激光共聚焦显微镜实验结果显示钙离子相对荧光(1.150 7±0.212 1)较加入CPA前显著增加(P<0.05)。加入抑制剂SKF-96365抑制SOCCs后,离体肌条收缩频率为(5.081 3±1.129 5)/min,较加入CPA后显著减缓(P<0.05),收缩幅度为(0.079 0±0.033 0)g,较加入CPA后显著增加(P<0.05);膜片钳显示钙电流较加入CPA后显著减少,-80 mV电流密度为(-2.678 1±0.804 4)pA/pF,20 mV电流密度为(0.907 7±0.346 2)pA/pF(P<0.05);激光共聚焦显微镜实验结果显示细胞内钙离子相对荧光(1.028 9±0.204 7)较加入CPA后显著减少(P<0.05)。结论膀胱平滑肌中可以表达SOCCs相关的通道,膀胱平滑肌的收缩性受到SOCCs的影响。Objective To investigate the function of intracellular store operated calcium channels （SOCCs） in the contraction of bladder smooth muscle of rats. Methods Bladder smooth muscle cells （BSMCs） and bladder smooth muscle strips were isolated from female SD rats. RT-PCR was employed to detect the expression of the SOCCs related genes in the bladder tissues. The SOCCs agonist CPA （10 μmol/L） and its inhibitor SKF-96365 （10 μmol/L） was used to treat the isolated BSMCs and muscle stripes. The isolated bladder smooth muscle was used for contraction in vitro. The cell current was recorded with whole-cell patch-clamp technique. The changes of intracellular calcium ion were observed by lacer scanning confocal microscopy. Results TRPC3, TRPC4 and STIM1 were expressed in BSMCs. CPA treatment resulted in significant increase in the contraction frequency （5.937 5±1.108 0 times/min） and obvious decrease in the amplitude （0.067 1±0.027 0 g） of the isolated muscle strips （P〈0.05）. Whole-cell patch-clamp technique indicated that CPA treatment induced obvious increase in the cell calcium currents, with the current destiny of -3.127 1±0.908 7 pA/pF at -80 mV and 1.048 1±0.322 5 pA/pF at 20 mV （P〈0.05）. The relative intensity of fluorescence was significant enhanced （1.150 7±0.212 1） than the cells without treatment （P〈0.05）. On the contrary, the use of SKF-96365 decreased the contraction frequency to 5.081 3±1.129 5 times/min and increased amplitude to 0.079 0±0.033 0 g of the bladder smooth muscle strips, both with significant difference with the addition of CPA （P〈0.05）. The calcium currents became little compared with that induced by CPA treatment, and the current destiny was -2.678 1±0.804 4 pA/pF at -80 mV and 0.907 7±0.346 2 pA/pF at 20 mV （P〈0.05）. The relative intensity of fluorescence was also significantly reduced （1.028 9±0.204 7） compared with the treatment of CPA （P〈0.05）. Conclusion The bladder smooth muscle expresses SOCCs, and its c

关 键 词：钙库调控的钙离子通道 膀胱平滑肌 钙离子 

分 类 号：R-332[医药卫生] R322.62