Protective effect of molecular hydrogen against H202-induced oxidative injury in cultured rat spinal cord neurons / 第二军医大学学报

ABSTRACT

Objective To investigate the protective effect of molecular hydrogen against hydrogen peroxide (H2O2)-induced oxidative injury in primary cultures of rat spinal cord neurons and the related mechanism. Methods Rat spinal cord neurons were cultured for 7 days and were randomly assigned to four groups with different treatments-(1) Normal medium (NM) control treated with NM; (2) Hydrogen-rich medium(HM) treated with HM; (3) NM+H2O2 treated with 100 μmol/L H2O2 and 15 μmol/L ferrous chloride (FeCl2) after pretreatment with NM for 2 h; and (4) HM+H2O2 treated with 100 μmol/L H2O2 and 15 μmol/L FeCl2 after pretreatment with HM for 2 h. The respective media were changed every 6 h in each group, and 12 h later 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-β. Results Compared with NM, HM significantly reduced the H2O2-induced intracellular production of ROS and HO • in purified neurons (P<0. 01), significantly decreased the number of apoptotic neurons (P < 0. 01) and expression of caspase-3 (P < 0. 01), and significantly promoted phosphorylation of GSK-β (P<0. 01). Conclusion Molecular hydrogen has protective effect against H2O2-induced oxidative injury in primary cultured neurons. Themechanisms might be related to reduction of intracellular production of ROS and HO •, inhibition of apoptosis in neurons and promotion of GSK-β phosphorylation.