ClC-3氯通道对Thapsigargin触发的Ca~(2+)运动的影响

目的探讨ClC-3氯通道与Thapsigargin(TG)触发的Ca2+运动的关系。方法在PC12细胞中转染全长ClC-3cDNA,利用生物荧光影像分析系统测定胞质Ca2+技术探讨ClC-3氯通道对TG触发的Ca2+运动的影响。结果与对照组相比,ClC-3蛋白过表达对TG触发的PC12细胞静息Ca2+]i的Ratio值和Ca2+释放的Ratio值无影响(P>0.05)。但使Ca2+内流量明显降低(P<0.05)。SK&F96365可以浓度依赖的抑制TG触发的Ca2+内流,但与对照细胞及空载体转染细胞相比,SK&F96365对ClC-3蛋白过表达细胞Ca2+内流的抑制作用明显减弱(P<0.05)。结论ClC-3蛋白参与TG触发的经Ca2+池操纵的Ca2+内流(store-operated Ca2+entry,SOCE)调节。

Effects of ClC-3 chloride channel on the Ca2+ movement induced by Thapsigargin in PC12 cells

Aim To investigate the relationship between ClC-3 chloride channels and Ca2+ movement induced by thapsigargin(TG)in PC12 cells.Methods The concentration of intracellular free calcium(Ca2+]i)expressed as the ratio value(Intensity340/Intensity380)was determined with Fura-2/AM probe.Results The overexpression of ClC-3 protein caused a significant decrease in TG-induced Ca2+ influx compared with control cells,and cells transfected with pcDNA3.1,whereas the Ca2+]i at resting level and at peak level was not different among all groups(P>0.05).SK & F96365 at the concentration of 5～20 μmol·L-1 inhibited the Ca2+ influx induced by 1.0 μmol·L-1 Thapsigargin in a concentration-dependent manner.The inhibitory effect of SK & F96365 on Ca2+ influx was decreased by overexpression of ClC-3 protein.Conclusion ClC-3 chloride channel was involved in the regulation of store-operated Ca2+ entry(SOCE).