目的:探讨硫化氢(H 2S)对缺氧诱导的皮层神经元损伤的影响及作用机制。方法:将SD大鼠皮层神经元在2% O 2、5% CO 2、93 % N 2、37 ℃培养箱培养24 h，建立细胞缺氧模型。以硫氢化钠（NaHS）作为H 2S的供体，应用CCK-8分析细胞活性；采用荧光探针DCFH-DA检测神经元活性氧(ROS)含量；用Rh123染色测定线粒体膜电位(MMP)；采用乳酸脱氢酶(LDH)试剂盒分析神经元LDH释放率，反映神经元的损伤情况。结果:(1)缺氧引起神经元ROS含量和LDH释放率升高，NaHS预处理可抑制缺氧所致神经元ROS含量和LDH释放率的升高；(2)缺氧降低神经元MMP和细胞活性，NaHS和活性氧清除剂NAC预处理均显著抑制缺氧所致神经元MMP和细胞活性的降低。结论:缺氧增加神经元ROS含量，降低神经元MMP和细胞活性，而H 2S通过其抗氧化作用，减轻缺氧所致神经元的损伤。
AIM:To explore the role of hydrogen sulfide (H 2S) in cortial neuronal injury induced by hypoxia.METHODS:The SD rat cortical neurons were cultured in hypoxic conditions (2% O 2, 5% CO 2 and 93% N 2 at 37 °C) to establish the hypoxic model. Sodium hydrosulfide (NaHS) was used as the donor of H 2S and neuronal viability was detected by CCK-8 assay. Neuronal content of reactive oxygen species (ROS) was determined by DCFH-DA method, and mitochondrial membrane potential (MMP) was detected using Rh123 staining. Lactate dehydrogenase (LDH) release rate was measured by a commercial kit to reflect the degree of neuronal injury. RESULTS:Hypoxic treatment increased ROS content and the release rate of LDH in the neurons. However, NaHS pretreatment significantly inhibited the hypoxia-induced increases in ROS content and LDH release. Hypoxia decreased MMP and cell viability. Pretreatment with NaHS and N-acetyl-L-cysteine (NAC), a ROS scavenger, significantly inhibited the decreases in MMP and viability of the neurons. CONCLUSION:Hypoxia induces ROS generation in the neurons, thereby decreases MMP and neuronal viability. H 2S significantly attenuates hypoxia-induced neuronal injury by its antioxygenation.
