中文摘要：

目的：探讨硫化氢（ H2 S）对氧糖剥夺/复氧（ OGD/R）诱导神经元损伤的影响。方法小鼠皮层神经元用无糖厄尔氏平衡盐溶液（EBSS）于2％ O2、5％ CO2、93％ N237℃培养4h，换用neurobasal ＋B27培养基于37℃5％CO2培养箱中继续培养12h，建立OGD/R模型。以硫氢化钠（NaHS）作为H2S的供体，采用细胞计数试剂盒（CCK-8）检测细胞存活率；用fura-2/AM和显微荧光成像系统检测神经元胞质钙离子浓度（[Ca^2＋]i）；利用比色法检测乳酸脱氢酶（ LDH）释放率；碘化丙啶（ PI）染色检测细胞损伤。结果200、300与600μmol/L NaHS预处理30min再OGD/R，神经元存活率显著提高（n＝4）；300μmol/L NaHS预处理后再OGD/R，[Ca^2＋]i（n＝5）、LDH释放率（ n＝4）和细胞损伤率（ n＝6）均低于OGD/R组，且使用10μmol/L钙螯合剂BAPTA也降低OGD/R诱导的LDH释放率和细胞损伤率。结论 H2 S减轻OGD/R所致皮层神经元损伤，其机制与H2 S抑制OGD/R诱导神经元钙超载有关。

英文摘要：

Objective To explore the effects of H 2 S on neuronal injuries induced by oxygen glucose deprivation /reoxygenation （ OGD/R） in cortical neurons .Methods For OGD, the primary cultured cortical neurons were incubated with glucose-free EBSS media for 4h in N2/CO2/O2 （93%/5%/2%） atmosphere.Thereafter, the media were replaced by Neurobasal/B27 culture media and the neurons were incubated for 12 h in a 5%CO2 incubator at 37℃.NaHS was used as a H2S donor and cell survival rate was determined by cell counting kit 8（CCk-8）.[Ca^2＋]i was determined using fura-2/AM and fluorescence microscopic imaging systems .The release rate of lactate dehydrogenase （ LDH） was determined by lactate dehydrogenase assay kit , and cell damage was analyzed by staining of propidium iodide （ PI ） .Results After pretreated with 200, 300 and 600μmol/L sodium hydrosulfide （ NaHS） for 30min before OGD/R, the cell survival rate of neurons significantly increased （n=4）.[Ca^2＋]I（n=5）, LDH release rate （n=4） and cell damage percentage （n=6） in the neuron pretreated with 300 μM NaHS were significantly lower than those in ODG/R cells.Treatment with 10μmol/L calcium chelator BAPTA also reduced the LDH release rate and cell damage percentage induced by ODG /R in neurons . Conclusion The results indicate that H 2 S may inhibit the OGD/R induced damage in cortical neurons via reducing calcium overload of neurons .