Effects Of Hydrogen On Mitochondrial Function And Neuronal Apoptosis In Rats With Cerebral Ischemia/Reperfusion Injury

Objective:To investigate the effects of hydrogen gas on mitochondrial function and neuronal apoptosis in the cortical region of rats with cerebral ischemia/reperfusion injury.Methods:A total of 78 male SD rats were selected in this experimental.The focal cerebral I/R injury model in rats was established by emboling the left middle cerebral artery for 2 hours and reperfusion for 24 hours.Rats were randomly divided into sham operation group(sham group)(n=26),brain I/R injury model group(Mod group)(n=26),hydrogen treatment group(H2 group)(n=26)by digital table method.After 24 hours of reperfusion,the neurological deficit score was performed;TTC staining was used to detect cerebral infarct size;The morphological changes of nerve cells were observed by Nissl ’s staining;Determination of reactive oxygen species in mitochondria of ischemic region by fluorescence spectrophotometry;The changes of Mitochondrial Membrane Potential(ψm)in Neurons was detected by JC-1 Staining;The opening degree of mitochondrial permeability transition pore(MPTP)were detected by a fluorescent enzyme labeling instrument;TUNEL staining were used to detect neuronal cell death and calculate the apoptotic index(AI);The expression of Bcl-2 and Bax was detected by Western blotting.Results:Compared with Sham group,the neurological deficit score,cerebral infarct size,mitochondrial ROS formation rate,the opening degree of MPTP,the AI and the Bax protein expression were significantly increased in Mod group(p<0.01),the mitochondrial membrane potential and the Bcl-2 protein expression were significantly decreased(p<0.01).Compared with Mod group,the neurological deficit score,cerebral infarct size(P<0.05)the mitochondrial ROS formation rate,the opening degree of MPTP,the AI(P<0.01)and the Bax protein expression(P<0.05)were significantly decreased in H2 group,the mitochondrial membrane potential and the Bcl-2 protein expression were significantly increased(P<0.01).Conclusion:Hydrogen has a protective effect on the mitochondrial function of cerebral ischemia reperfusion injury.The mechanism may be related to decreasing the production rate of mitochondrial reactive oxygen species,maintaining the mitochondrial membrane potential level of neurons,and decreasing mitochondrial permeability transition pore opening after I/R damage.On the other hand,hydrogen can reduce the apoptosis of neural cells in mitochondrial pathway,reduce the area of cerebral infarction in rats and improve the neurological function score after I/R in the rat brain.The mechanism may be related to the up-regulation of the expression of Bcl-2,the down-regulation of the expression of Bax and the decrease of MPTP opening.