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目的:研究皮质酮(Cor)对原代培养海马神经细胞存活和海马神经细胞电压依赖性钙通道(VDCC)的影响。方法:原代海马神经细胞存活率测定用MTT比色法。海马神经细胞上VDCC内向Ca~(2+)电流检测采用全细胞膜片箝技术。结果:Cor可浓度依赖地损伤原代海马神经细胞和皮层神经细胞,IC_(50)分别为3.2μmol·L~(-1)和85μmol·L~(-1),Cor(1μmol·L~(-1)-0.1mmol·L~(-1))喷射于海马神经细胞表面即刻显著促进电压依赖性Ca~(2+)内流,其最大升幅分别是53％,191％和84％,而且Cor诱导的钙内流增加是非浓度依赖和非电压依赖的。结论:Cor可显著促进海马神经细胞电压依赖性钙通道开放,该作用可能是Cor海马神经毒性作用的机制之一。
Objective: To investigate the effect of cortisone (Cor) on the survival of primary cultured hippocampal neurons and the voltage-dependent calcium channel (VDCC) in hippocampal neurons. Methods: Primary hippocampal neuronal cell viability was assayed by MTT colorimetry. The whole cell patch clamp technique was used to detect the inward Ca 2+ current in VDCC of hippocampal neurons. RESULTS: Cor could damage the primary cultured hippocampal neurons and cortical neurons in a concentration-dependent manner. IC 50 was 3.2 μmol·L -1 and 85 μmol·L -1, Cor 1 μmol·L -1 -1) -0.1 mmol·L -1) significantly accelerated voltage-dependent Ca 2+ influx at the surface of hippocampal neurons, with the largest increases being 53%, 191% and 84%, respectively, and Cor-induced increases in calcium influx were non-concentration-dependent and voltage-independent. Conclusion: Cor can significantly promote the voltage-dependent calcium channel opening in hippocampal neurons, which may be one of the mechanisms of neurotoxicity in Cor hippocampus.