Protective effects of hydrogen-rich water postconditioning on glutamate injury of brain slices of neonatal rats / 中国中西医结合急救杂志

ABSTRACT

Objective To investigate the protective effects of hydrogen-rich water postconditioning on glutamate (Glu) mediated ischemia/reperfusion (I/R) injury in isolated brain slices of neonatal rats and explore its mechanism of action.Methods The brains of Sprague-Dawley (SD) neonatal rats aged 7 days were cut into slices and cultured. And then the cultured slices were randomly divided into the normal control group, Glu injury group (1 mmol/L Glu for 30-minute injury), hydrogen-rich water postconditioning group (after Glu injury for 30 minutes, cultured with complete medium containing 100μmol/L of hydrogen-rich water), once per 3 hours to change the medium for totally 24 hours, each group having 12 holes. The brain slices were stained by hematoxylin-eosin (HE) staining to observe the changes of nerve cells. The lactate dehydrogenase (LDH) release rates, the numbers of nissl bodies and the basic fibroblast growth factor (bFGF) in each brain slice were determined to evaluate the degree of cerebral neuronal damage.Results Compared with the normal control group, the number of nerve cells was rare, and the structure not complete; the LDH release rate and the number of bFGF were increased significantly in Glu injury groups [LDH release rates (50.66±4.93)% vs. (20.15±5.14)%, bFGF (cells/400 power field) 22.79±2.13 vs. 4.13±1.17, both P < 0.01); the Nissl body was decreased (cells/400 power field 11.81±2.69 vs. 47.10±3.78,P < 0.01) in Glu injury group. Compared with Glu injury group, the morphological structure of brain nerve cells was restored, the LDH release rate was reduced [(39.13±3.66)% vs. (50.66±4.93)%]; bFGF was decreased (cells/400 power field 14.22±1.22 vs. 22.79±2.13), and the Nissl body was increased (cells/400 power field 23.25±6.05 vs. 11.81±2.69) in hydrogen-rich water postconditioning group (allP < 0.05).Conclusions Hydrogen-rich water postconditioning has protective effects on rat brain slices with I/R injury induced by Glu. Its mechanism was possibly related to the reduction of free radicals, calcium overload and inflammatory factors induced by excitatory amino acids toxicity, resulting in inhibition of cell apoptosis.