蝙蝠葛苏林碱抑制NMDA引起的细胞游离钙升高而减少神经毒性

Daurisoline reduced neurotoxicity by inhibiting NMDA induced intracellular free calcium level

目的：研究蝙蝠葛苏林碱（ｄａｕｒｉｓｏｌｉｎｅ，ＤＡＵ） 拮抗Ｎ甲基Ｄ精氨酸（ ＮｍｅｔｈｙｌＤａｓｐｒａｔａｔｅ，ＮＭＤＡ） 引起的神经毒性作用，并对其作用机制进行了分析。方法：ＮＭＤＡ 损伤原代培养的海马神经细胞；培养细胞内游离钙浓度的测定；大鼠动脉条内８６ Ｒｂ＋ 外流率的测定。结果：ＤＡＵ 能对ＮＭＤＡ 损伤的原代培养的海马神经细胞有明显的保护作用，能剂量依赖性的抑制损伤后培养细胞中ＬＤＨ 的释放；ＤＡＵ 还能剂量依赖性的抑制ＫＣｌ，Ｂａｙ，Ｋ８６４４，ＮＭＤＡ，去甲肾上腺素（ＮＥ） 以及咖啡因（ｃａｆｆｅｉｎｅ） 刺激的细胞内游离钙浓度的上升，但并不影响静息的或由钾通道开放剂吡哪地尔（ｐｉｎａｃｉｄｉｌ，ＰＩＮ） 刺激的大鼠动脉条内８６ Ｒｂ＋ 的外流率。结论：ＤＡＵ 通过阻断ＮＭＤＡ 引起的细胞钙升高而拮抗神经毒性。

OBJECTIVE:To study the effect of daurisoline (DAU) against on N methyl D aspratate (NMDA) induced injury and the effects of DAU on calcium and potassium channel.METHODS:Primary cultured hippocampus neurons were injured by NMDA and the LDH levels in culture medium were tested as cell availability;intracellular free calcium level in PC12 and NG108 cells were tested by Fura 2 method;and 86 Rb + efflux from rat aorta were examined for potassium channel behavior.RESULTS:The data showed that NMDA (100 μmol·L -1 ) induced widespread cell injury which was substantially attenuated by adding of DAU (0.01～1μmol·L -1 ).DAU inhibited NMDA stimulated LDH release in the culture in a dose dependent manner.NMDA evaluated the intracellular free calcium ihn PC 12 cells were also blocked by DAU in a dose dependent manner with its IC 50 volume of 6 36μmol·L -1 .DAU had no effect on spontaneous or pinacidil,a potassium channel opener,stimulated 86 Rb + efflux.CONCLUSION:The results indicated that NMDA induced neurotoxicity could be blocked by DAU with its ability of inhibiting calcium channel directly.