CAJ | 학술논문

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（[Ca2＋]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，[Ca2＋]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。
목적：탐토류화경（ H2 S）대양당박탈/복양（ OGD/R）유도신경원손상적영향。방법소서피층신경원용무당액이씨평형염용액（EBSS）우2％ O2、5％ CO2、93％ N237℃배양4h，환용neurobasal ＋B27배양기우37℃5％CO2배양상중계속배양12h，건립OGD/R모형。이류경화납（NaHS）작위H2S적공체，채용세포계수시제합（CCK-8）검측세포존활솔；용fura-2/AM화현미형광성상계통검측신경원포질개리자농도（[Ca2＋]i）；이용비색법검측유산탈경매（ LDH）석방솔；전화병정（ PI）염색검측세포손상。결과200、300여600μmol/L NaHS예처리30min재OGD/R，신경원존활솔현저제고（n＝4）；300μmol/L NaHS예처리후재OGD/R，[Ca2＋]i（n＝5）、LDH석방솔（ n＝4）화세포손상솔（ n＝6）균저우OGD/R조，차사용10μmol/L개오합제BAPTA야강저OGD/R유도적LDH석방솔화세포손상솔。결론 H2 S감경OGD/R소치피층신경원손상，기궤제여H2 S억제OGD/R유도신경원개초재유관。
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).[Ca2+]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).[Ca2+]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 .