ATP激活雪旺细胞内游离Ca^(2+)浓度机制的实验研究

Increase of intracellular Ca^(2+) concentration by extracellular ATP: in vitro study of its mechanism

目的 探讨细胞外ATP引起的细胞内游离Ca2 +浓度升高的机制。方法 根据细胞内游离Ca2 + 的来源 ,分别在加入细胞外ATP的雪旺细胞的培养基中加入P2Y受体阻滞剂苏拉明和Ca2 + 通道阻滞剂异博定作为实验组 ;同时 ,以仅加入细胞外ATP的雪旺细胞的培养基作为对照组。实验组和对照组均使用Fura 2染色剂进行染色。结果 使用P2Y受体阻滞剂苏拉明的雪旺细胞的光反射强度减弱 ;而使用Ca2 + 通道阻滞剂异博定的雪旺细胞的光反射强度基本不变。结论 细胞外ATP可能是通过P2Y受体介导激活磷脂酶C引起细胞内IP3 增加 ,使Ca2 + 自储库中释放 ,导致细胞内游离Ca2 + 浓度升高。

ObjectiveTo investigate the mechanism of the rise of intracellular Ca 2+ concentration caused by extracellular ATP.MethodsSchwann cell isolation cultured with 100 uM ATP was supplimented with 2 different agents. In one group, Suramin, a P 2Y receptor blocker, was added into the culture media. In the other group, Verapamil, a calcium channel blocker, was added. Another group of Schwann cell culture supplimented with ATP only served as control. After 48 hours combined culture, supernatant of each group was stained by Fura 2 to detect intracellular Ca 2+ concentration of the Schwann cells.ResultsThe fluorescent density of the supernatant in the culture supplimented with Suramin was attenuated, whileas that of the supernatant of the culture supplimented with Verapamil was not changed as compared to the control.ConclusionExtracellular ATP could activate phospholipase C via P 2Y receptor which causes the elevation of IP 3 concentration. IP 3 in turn would lead to release of Ca 2+ from endoplasmic reticulum and result in rise of intracellular Ca 2+ concentration.