反义ClC-3寡核苷酸对thapsigargin触发的Ca^2＋运动的影响  

Effects of ClC-3 Antisense Oligonucleotide on Ca^(2+) Movement Induced by Thapsigargin in PC12 Cells

作　　者：张海宁[1] 丘钦英[1] 关永源[1] 

机构地区：[1]中山大学中山医学院药理学教研室

出　　处：《中山大学学报（医学科学版）》2008年第5期526-530,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(30271503);中华医学会基金(CMB,00730);国家教育部博士点基金(20020558055);广东省自然科学基金团队项目(5300822)

摘　　要：【目的】研究反义ClC-3寡核苷酸对Thapsigargin(TG)触发的Ca2+运动的影响。【方法】在PC12细胞中转染反义ClC-3寡核苷酸,利用生物荧光影像分析系统测定胞浆Ca2+技术探讨ClC-3寡核苷酸对TG触发的Ca2+运动的影响。【结果】与对照组相比,反义寡核苷酸转染对TG触发的PC12细胞静息[Ca2+]i的Ratio值和Ca2+释放量的Ratio值无显著影响(P>0.05)。但使Ca2+内流量明显升高(P<0.05)。Ca2+池操纵性Ca2+通道(store-operatedCa2+channels,SOCC)阻断剂SK&F96365可以浓度依赖的抑制TG触发的PC12细胞Ca2+内流,但与随义、正义寡核苷酸转染组相比,SK&F96365对反义转染组细胞Ca2+内流的抑制作用明显增强(P<0.05)。【结论】ClC-3蛋白参与TG触发的Ca2+池操纵的Ca2+内流(store-operatedCa2+entry,SOCE),但对细胞静息钙水平及钙释放过程没有影响。[Objective] To investigate the effects of ClC-3 chloride channels on the Ca^2＋ movement induced by thapsigargin （TG） in PC12 cells transfected with ClC-3 oligonucleotides. [Methods] The concentration of intracellular free calcium （[Ca^2＋]i） expressed as the ratio value （Intensity340/ Intensity380） was determined with Fura-2/AM probe. [Results] The results showed that transient transfection of PC12 cells with antisense oligonucleotide specific to ClC-3 caused a significant increase in TG-induced Ca^2＋ influx compared with that in control cells, and cells transfected with lipofectamine, missense or sense oligonucleotide, whereas the [Ca^2＋]i at resting level and at peak level were not different among all groups （P〉0.05）. SK＆F96365, a blocker for storeoperated calcium channels （SOCC）, inhibited the Ca^2＋ influx induced by 1.0 μmol/L thapsigargin in PC12 cells in a concentration-dependent manner. The inhibitory effect of SK＆F96365 on Ca^2＋ influx was enhanced by antisense oligonucleotide as compared with Lipofectamine, sense or missense oligonuclotide. [Conclusion] C1C-3 chloride channels was involved in TG-induced store-operated Ca^2＋ entry（SOCE）, but had no effects on the [Ca^2＋]i at resting level and Ca^2＋ release from Ca^2＋ stores.

关 键 词：反义ClC-3寡核苷酸 钙离子 THAPSIGARGIN 氯通道 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]