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目的探讨缺氧条件下阻断酸敏感离子通道1α(acid sensing ion channels 1α,ASIC1α)对大鼠皮质神经元存活及细胞内钙离子水平的影响,以及缺氧对皮质神经元ASIC1表达的影响。方法通过使用ASIC1α阻断剂、N甲基D门冬氨酸(NMDA)受体阻断剂、电压门控通道阻断剂,利用fura-3/AM荧光显像,乳酸脱氢酶(lactate dehydrogenase,LDH)释放实验等技术,比较观察对缺氧皮质神经元存活以及钙离子水平的影响,进一步利用Western印迹法检测ASIC1的表达变化。结果酸性环境中,缺氧皮质神经元形态明显改变,细胞内钙离子浓度升高;与加入NMDA受体阻断剂、电压门控通道阻断剂相比较,加入ASIC1α阻断剂明显抑制细胞内钙离子水平升高(P<0.05),对神经元起到保护作用。随缺氧时间的延长,神经元ASIC1蛋白表达增加。结论酸性环境中,缺氧皮质神经元的损伤与ASIC1α开放以及表达增加有关;阻断ASIC1α具有神经保护作用。
Objective To investigate the effects of inhibiting acid sensing ion channels 1α (ASIC1α) on the survival of cortical neurons and the level of intracellular Ca2 + in hypoxia and the effect of hypoxia on the expression of ASIC1 in cortical neurons. Methods By using ASIC1α blockers, NMDA receptor blockers, voltage-gated channel blockers, fura-3 / AM fluorescence imaging, lactate dehydrogenase , LDH release experiments and other techniques were compared to observe the survival of hypoxic cortical neurons and calcium levels, and further using Western blot assay ASIC1 expression changes. Results In acidic environment, the morphology of hypoxic cortical neurons changed obviously and the concentration of intracellular calcium increased. Compared with the addition of NMDA receptor blocker and voltage gated channel blocker, the addition of ASIC1α inhibitor significantly inhibited intracellular The level of calcium increased (P <0.05), which protected the neurons. With hypoxia time, neuronal ASIC1 protein expression increased. Conclusion In acidic environment, the injury of hypoxic cortical neurons is related to the opening of ASIC1α and the increase of the expression of ASIC1α. Blocking ASIC1α has the neuroprotective effect.