硫化氢对缺氧诱导的大鼠皮层神经元损伤的保护作用  

Protective effect of hydrogen sulfide on hypoxia-induced injury of rat cortical neurons

作　　者：魏楚蓉[1] 刘路宽[1] 田立兵 余艳贞 曾九江[1] 白帅[1] 毛慕华[1] 文秀华[1] 罗友根[1] 

机构地区：[1]井冈山大学医学院神经退行性疾病与衰老研究中心,江西吉安343000

出　　处：《中国病理生理杂志》2014年第2期203-207,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31360238);江西省自然科学基金资助项目(No.20132BAB205020);井冈山大学2013年度国家级大学生创新创业训练计划资助项目(No.2013001);吉安市科技支撑资助项目[No.(2012)32号10];井冈山大学自然科学科研项目(No.JZ1210)

摘　　要：目的:探讨硫化氢(H2S)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%O2、5%CO2、93%N2、37℃培养箱培养24 h,建立细胞缺氧模型。以硫氢化钠(NaHS)作为H2S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H2S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 （ H2 S） in cortial neuronal injury induced by hypoxia. METHODS： The SD rat cortical neurons were cultured in hypoxic conditions （2% O2, 5% CO2 and 93% N2 at 37 ℃） to establish the hypoxic model. Sodium hydrosulfide （NariS） was used as the donor of H2S and neuronal viability was de- tected by CCK-8 assay. Neuronal content of reactive oxygen species （ROS） was determined by DCFH-DA method, and mi- tochondrial 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, NariS pretreatment significantly inhibited the hypoxia-in- duced increases in ROS content and LDH release. Hypoxia decreased MMP and cell viability. Pretreatment with NariS 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. H2 S significantly attenuates hypoxia-induced neuronal injury by its antioxygenation.

关 键 词：缺氧 神经元损伤 硫化氢 活性氧类 

分 类 号：Q421[生物学—神经生物学]