CNQX对乙醇、乙醛调控大脑皮质神经元神经甾体水平的影响

目的 研究乙醇、乙醛对原代培养大鼠大脑皮质神经元神经甾体水平的影响以及与AMPA受体调控的相关性. 方法 应用AMPA受体拮抗剂CNQX阻断AMPA受体,用50mmol/L乙醇或25 mmol/L乙醛处理神经元,收集细胞培养上清,采用高效液相色谱质谱法分离测定游离型神经甾体孕烯醇酮(PREG)、脱氢表雄酮(DHEA)、别孕烯醇酮(AP)及结合型神经甾体脱氢表雄酮硫酸酯(DHEAS)、孕烯醇酮硫酸酯(PREGS)的含量,观察CNQX对乙醇、乙醛调控原代培养神经元神经甾体水平的影响. 结果乙醇可明显升高神经元PREG、AP和PREGS的水平,与PBS对照组比较差异有统计学意义(P＜0.05);乙醛可明显升高神经元PREG、DHEA、PREGS和DHEAS的水平,与PBS对照组比较差异有统计学意义(P＜0.05).CNQX反转了乙醇升高PREG、PREGS水平的作用以及乙醛升高PREG、DHEAS水平的作用,却使AP水平明显升高,组间比较差异有统计学意义(P＜0.05). 结论 乙醇、乙醛对大脑皮质神经元神经甾体PREG、DHEA、AP、PREGS和DHEAS的影响各不相同,CNQX反转了乙醇上调PREG、PREGS水平的作用以及乙醛上调PREG、DHEAS水平的作用.

CNQX affecting regulation abilities of ethanol and aldehyde on levels of neurosteroid in cortical neurons

Objective To investigate the effects of ethanol and aldehyde on the levels of neurosteroid in the cortical neurons of primary-cultured rots and explore their relations with the regulation of AMPA receptor. Methods CNQX (the AMPA receptor antagonist) was used to block the AMPA receptor, and then, 50 mmol/L ethanol or 25 mmol/L aldehyde was employed to treat the neurons;supernate was collected. HPLC-MS/APCI was employed to analyze and determine the concentrations of unconjugated neurosteroids (pregnenolone PREG], dehydroepiandrosterone DHEA], allopregnanolone AP]) and conjugated neurosteroids (dehydroepiandrosterone DHEAS], pregnenolone sulphate PREGS]) in media from primary cultural cerebral cortex neurons. The effects of CNQX on the regulation abilities of ethanol and aldehyde on levels of neurosteroids in primary-cultured cerebral cortex neurons were detected. Results The levels of PREG, AP and PREGS in the ethanol treatment group were significantly increased as compared with those in the PBS treatment group (P＜0.05); the levels of PREG,DHEA,PREGS and DHEAS in the aldehyde treatment group were significantly increased as compared with those in the PBS treatment group (P＜0.05). CNQX reversed the effect of ethanol on increasing the levels of PREG and PREGS, and reversed that of aldehyde on increasing the levels of PREG and DHEAS, but induced the effect on increasing the level of AP; these effects were significantly different between each 2 groups (P＜0.05). Conclusion Both ethanol and aldehyde show different effects on levels ofneurosteroids in primary-cultured cerebral cortex neurons. CNQX reverses the effect of ethanol on increasing the levels of PREG and PREGS, and that of aldehyde on increasing the levels of PREG and DHEAS.