| 摘要: |
| 目的 证明氢分子对过氧化氢（H2O2）诱导氧化损伤的大鼠脊髓神经元具有保护效应，并初步探讨相关机制。方法 将纯化培养7天后的大鼠脊髓神经元分为四组：（1）常规培养基（NM）对照组：以NM继续培养；（2）富氢培养基（HM）组：以HM继续培养；（3）NM + H2O2组：以NM培养2h后，加入100 μM H2O2和15 μM FeCl2继续培养；（4）HM + H2O2组：以HM预处理2h后，加入100 μM H2O2和15 μM FeCl2继续培养。各组处理后每6小时更换各自培养基及氧化剂，在12h后停止处理并检测神经元胞内氧自由基生成的水平、凋亡情况，以及GSK-3β和p-GSK-3β的表达水平。结果 与普通培养基相比，富氢培养基可降低H2O2氧化损伤后神经元胞内ROS尤其是HO•的生成（P ＜ 0.01）、减少神经元凋亡的数量（P ＜ 0.01）并下调Caspase-3的表达（P ＜ 0.01），以及促进GSK-3β的磷酸化（P ＜ 0.01）。结论 氢分子对H2O2氧化损伤的神经元具有保护效应，其机制与降低神经元胞内ROS尤其是HO•的生成、减少神经元凋亡的数量和抑制凋亡信号通路的激活、以及促进GSK-3β的磷酸化利于神经元生长有关。 |
| 关键词: 氧化损伤 过氧化氢 氧自由基 氢分子 神经元 凋亡 糖原合成激酶-3β |
| DOI：10.3724/SP.J.1008.2014.00233 |
| 投稿时间：2013-09-25修订日期：2013-12-19 |
| 基金项目:全军医学科技“十二五”科研项目（CWS11J121）；国家自然科学基金资助项目（81200953） |
|
| Protective effect of molecule hydrogen on H2O2-induced oxidative injury in cultured rat spinal cord neurons |
| liu fang ting,li jian,li xiang nan,xu sheng ming,xiang zheng hua,yuan hong bin |
| (Department of Anesthesiology,Changzheng Hospital, Second Military Medical University;Department of Orthopaedics, Changzheng Hospital, Second Military Medical University;Department of Neurobiology, Second Military Medical University) |
| Abstract: |
| Objective To investigate the protective effect of molecular hydrogen against hydrogen peroxide（H2O2）-induced oxidative injury and the correlated mechanism in primary cultures of rat spinal cord neurons. Methods Primary cultures of rat spinal cord neurons were cultured for 7 days, and then were randomly assigned to four groups with different treatments. (1) Normal medium (NM) control: continued to culture with normal medium. (2) Hydrogen-rich medium（HM）: continued to culture with hydrogen-riched medium. (3) NM + H2O2: cultured with NM, and treated with 100 μM H2O2 and 15 μM ferrous chloride (FeCl2) after pretreatment of HM for 2 h. (4) HM + H2O2: cultured with HM containing 100 μM H2O2 and 15 μM FeCl2 after pretreatment of HM for 2 h. The respective media were changed every 6 h. Treatments were carried out for 12 h, and then the cells were collected for assays of reactive oxygen species (ROS), hydroxyl radical (HO•), apoptosis, and the expression of Glycogen synthase kinase-3β (GSK-3β) and p-GSK-3β. Results Compared with NM, HM could reduce the H2O2-induced intracellular production of ROS and HO• in purified neurons (P ＜ 0.01), and the number of apoptotic neurons was significantly decreased (P ＜ 0.01) with the downregulation of Caspase-3. In addition, HM also promoted the phosphorylation of GSK-3β (P ＜ 0.01). Conclusion Molecular hydrogen possessed the protective effect against H2O2-induced oxidative injury in primary cultured neurons. The mechanisms were related to reducing the intracellular production of ROS and HO•, inhibiting the activation of apoptosis in neurons and increasing the phosphorylation of GSK-3β to promote growth of neurons. |
| Key words: oxidative injury H2O2 ROS molecular hydrogen neurons apoptosis GSK-3β |
