Dynamic Expression Of Bcl-2, Bax, And Caspase-3 In Ischemia-reperfusion Brain Cortex Of Rat

Objective A lot of studies have shown that cerebral ischemia and reperfusion in neuronal death, including necrosis and apoptosis, there is also neuronal regeneration and repair, participating in a series of apoptosis in regulating apoptosis in Bcl-2, Bax and Caspase-3 gene is an important apoptosis genes, which may also neuronal regeneration and repair play an important role. This study observed after ischemia and reperfusion 2h,3h,6h,8h,12h of Bcl-2, Bax and Caspase-3 protein expression to investigate ischemia-reperfusion apoptosis and Bcl-2, Bax and Caspase-3 protein expression, analysis its possible role in neurogenesis and repairing.Method Clean grade healthy male (Sprague-Dawley) rats were 66, weight 250±20 g, were randomly divided into ischemia reperfusion 2h,3h,6h,8h,12h group and sham operation control group, n=12 animals. Experimental group were caused by using the intraluminal suture method of middle cerebral artery occlusion (Middle Cerebral Artery Occlusion, MCAO),2h reperfusion, according to reperfusion time points were divided into 2h group,3h group,6h group,8h group,12h group. With 4% paraformaldehyde by cardiac perfusion, and brain slices in situ detection of TUNEL after row of apoptosis and Bcl-2, Bax and Caspase-3 immunohistochemical staining. All results are mean±standard deviation (x±s) that SPSS 10.0 statistical software used to analyze experimental data, p<0.05 for the difference was significant.Results The TUNEL staining showed rare in the sham operation group neuronal apoptosis and cerebral ischemia and ischemic reperfusion groups around the area of apoptotic cells in the 2,6 and 12 hours gradually increased. Adjacent to the time focal cerebral ischemia and reperfusion apoptosis and Bcl-2 and Bax gene expression point of comparison, significant difference (P<0.05). Sham group, Bcl-2 protein have a certain expression of cerebral ischemia and reperfusion in each group increased expression of Bcl-2 protein,8h,12h express the most obvious. Adjacent to each time point, in addition to 8h,12h group, the expression of differences among the groups were significantly (p<0.05). Sham group, a small amount of Bax protein expressionCerebral ischemia and reperfusion groups 2h after reperfusion increased gradually to 12 hours, the most obvious expression of adjacent each group, except the sham operation group and reperfusion 2h group, the difference was significant (P <0.05). Sham group rat brain in the expression of Caspase-3 are weak; 2 hours after ischemia, ischemic side of the Caspase-3 positive cells was no significant increase in 3 hours ischemic side Caspase-3 positive cells began to increase,6 hours ischemic side Caspase-3 positive cells significantly increased the 2010 draft papers, 8 hours and 12 hours further increased. Adjacent to each group, except the sham operation group and reperfusion 2h group, the difference was significant (P<0.05).Conclusion 1. Bcl-2, bax protein expression with ischemia time as an increasing tendency. Bcl-2 expression in cell morphology is relatively complete, but bax expression in cells severely damaged.2. Caspase-3 protein expression with ischemia time as an increasing tendency.