论文部分内容阅读
目的 研究ATP触发牛主动脉内皮细胞Ca2 + 内流特性 ,Cl-通道阻断剂furosemide和PKC抑制剂staurosporine对其影响 ,阐明Cl-通道和PKC与ATP触发Ca2 + 内流的内在联系。方法 采用Fura 2荧光测定胞浆Ca2 + 变化技术 ,在培养的乳牛主动脉内皮细胞上观察药物对ATP触发Ca2 + 内流的影响。结果 ATP触发的Ca2 + 内流不受电压依赖性钙通道 (VDC)阻断剂nifedipine影响 ,但可被非VDC阻断剂SK&F 96 36 5 ,非选择性阻断Ca2 + 通道的金属离子NiSO4 和PKC抑制剂staurosporine抑制。furosemide (1 2 5～ 10 μmol·L-1)呈浓度依赖性抑制ATP触发的Ca2 + 内流 ,10 μmol·L-1可抑制 36 %± 14%。当staurosporine(10 0nmol·L-1)或SK&F 96 36 5 (15 μmol·L-1)分别对Ca2 + 内流最大程度抑制 34 %± 5 %和 6 4%± 11%后 ,furosemide可分别进一步抑制 7 6 %± 4%和 14%± 7%。结论 furosemide敏感的Ca2 + 内流与SK&F 96 36 5敏感的Ca2 + 内流存在非同一性。furosemide敏感的Cl-通道开放与PKC激活均参与了ATP触发的Ca2 + 内流 ,这两种因素与Ca2 + 内流之间存在内在联系。
Objective To investigate the effects of ATP on Ca2 + influx in bovine aortic endothelial cells, the effects of furosemide, a Cl-channel blocker, and staurosporine, a PKC inhibitor, on the intrinsic relationship between Cl-channel, PKC and ATP-triggered Ca2 + influx. Methods The changes of Ca2 + in cytoplasm were detected by Fura2 fluorescence assay. The effects of drugs on Ca2 + influx induced by ATP were observed on cultured aortic endothelial cells. Results ATP-triggered Ca 2+ influx was not affected by nifedipine, a voltage-dependent calcium channel (VDC) blocker, but could be blocked by non-VDC blocker SK & F 96 36 5, non-selective blocking of CaSO4 + PKC inhibitor staurosporine inhibition. Furosemide (1 25 ~ 10 μmol·L-1) inhibited ATP-induced Ca2 + influx in a concentration-dependent manner, while 10 μmol·L-1 inhibited 36% ± 14% of the Ca2 + influx. Furosemide may be further further reduced when staurosporine (10 0 nmol·L-1) or SK & F 96 36 5 (15 μmol·L-1) maximally inhibits Ca2 + influx by 34% ± 5% and 6.4% ± 11%, respectively Inhibition of 7 6% ± 4% and 14% ± 7%. Conclusion The furosemide-sensitive Ca2 + influx is not identical to the SK2-infused Ca2 + influx. Both furosemide-sensitive Cl-channel opening and PKC activation are involved in ATP-triggered Ca2 + influx, an intrinsic relationship between Ca2 + influx and these two factors.