氧自由基对血管内皮细胞钙电流的影响及山莨菪碱的干预作用

Effect of OFR on the currents of store-operated Ca^(2+) channels in human vein endothelial cell and the intervention of Anisodamine

目的观察山莨菪碱(anisodamine,Ani)在氧自由基(oxygen free radical,OFR)作用体外培养人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)后对其钙池操纵的Ca2+通道电流(store-operated Ca2+currents,Isoc)的影响。方法利用全细胞膜片钳记录技术,应用快速加药灌流装置将0.5、1.0、1.5 mmol/L三种不同浓度的Ani对OFR作用HUVECs后ISOC的影响。结果OFR能增强HUVECs的ISOC,而Ani对OFR作用HUVECs后所引起ISOC的增大作用具有抑制效应。当膜电位钳制于-100 mV时,1.5 mmol/L Ani可使ISOC的峰值电流从(-388.59±58.66)pA下降至(-287.37±23.03)pA,抑制率为(25.013±9.49)%;OFR作用后,1.5 mmol/LAni可使ISOC的峰值电流从(-577.04±93.69)pA下降至(-302.38±35.13)pA,抑制率为(47.24±3.82)%。各组间及与对照组相比均有显著的统计学差异(P<0.01,n=6)。结论Ani具有明显抑制HUVECs细胞ISOC的作用,对OFR所致的血管内皮损伤确有保护作用。

Objective To study the effect of oxygen free radical （OFR） on the currents of store-operated Ca^2＋ channels （Isoc ）and the inhibitory effects of Anisodamine （Ani）on OFR-induced Isoc potentiation in human umbilical vein endothelial cells （ HUVECs ） cultured in vitro. Methods HUVECs were respectively stimulated by 0. 5,1. 0 and 1. 5 mmol/L levels of Ani as terminal concentration conditions after OFR stimulation in our experiments. The dynamic changes of Isoc of HUVECs were determined by tight-seal whole- cell patch-clamp techniques. Results OFR could potentialize Isoc of HUVECs, alternatively, Ani could inhibit this effect in a concentration-dependent manner. At the potential of - 100 mV, Ani inhibited the peak amplitude OfIsoc from （ -388.59± 58.66）pA to （ -287.37± 23.03） pA（P〈0.01,n=6）,with the inhibition rate of （25. 013±9.49）% at 1.5 mmol/L levels. After stimulation with 0.1 mmol/L OFR, the peak amplitude of Isoc in the HUVECs decreased by Ani and from （ -577.04 ± 93.69）pA to （ -302.38±35.13 ） pA, with the inhibition rate of （ 47.24 ±3.82 ） % at 1.5 mmol/L levels, without affecting their reversal potential of Isoc occurred in all cells. There was significant differences among these groups. Conclusion It is concluded that Anisodamine obviously suppress OFR-induced Isoc elevation which probably is one of the mechanisms of its protective effects on vascular endothelial cells.