Effect Of Mitochondrial Permeability Transition Pore On Mitochondrial Fission And Its Mechanism In Cerebral Ischemia Reperfusion Injury

Objective: To evaluate the effect of the mitochondrial permeability transition pore suffering from anoxia-reoxygenation injury and mechanismsMethods: Newborn Wistar rats within 24 h after birth were chosen as providers of primary cultured hippocampal neurons.Cells were random divided into 5 groups(n = 45).The normal group(C group): cells without any processing;Anoxia-reoxygenation group(I group)cells were subjected to 6h anoxia,and followed by 20hreoxygenation;Anoxia-reoxygenation and vehicle group(V group): cells were treated with DMSO,final concentration of DMSO <0.01%)for 40 min prior to anoxia;Anoxia-reoxygenation and ATR group(M group)cells were treated with 400 umol/L ATR(dissolve in DMSO,final concentration of DMSO <0.01%)for 40 min prior to anoxia;Anoxia-reoxygenation and CSA group(N group)cells were treated with 0.2 umol/L CSA(dissolve in DMSO,final concentration of DMSO <0.01%)for 40 min prior to anoxia.Enzyme standard instrumentwas used to examined the levels of ATP.western blotting was used to measure the expression of proteins Drp1,Cyt c and Bcl-2.Electron microscope and fluorescent microscopy were used to observe the Mitochondrial morphology and the levels of Mitochondrial membrane potential(MMP),and flow cytometer was used to examine the ratio of cell apoptosis.Results: The results showed that compared with group C,the mitochondria were significantly swollen,the matrix was evacuated,the inner and outer membrane separation,the rupture of the rupture,the destruction of the network structure,the ATP content and the mitochondrial membrane potential were significantly decreased,the apoptosis rate increased,Drp1 and Cyt c protein expression increased,while the expression of Bcl-2 decreased,the difference was statistically significant(P <0.05).Compared with group I,there was no significant difference in ATP content and protein expression between the two groups(P> 0.05).Compared with group I,the mitochondria in group N were slightly swollen,the matrix was compact and orderly.The net structure was completely preserved,the ATP content and mitochondrial membrane potential were significantly increased,the apoptotic rate was significantly decreased,the expression of Drp1 and Cyt c protein decreased,And the expression of Bcl-2 was significantly increased,the difference was statistically significant(P <0.05).And the experimental results of group M were opposite to those of N group.Conclusion: By inhibiting the mitochondrial permeability transition pore opening can obviously improve the cerebral ischemia-reperfusion injury in the mitochondrial division,its mechanism may be increased mitochondrial membrane potential.