Influence of Sevoflurane Post-processing on Bcl-2and Caspase-3Expression in Hippocampus of Cerebral Ischemia Reperfusion Rat Model
|Keywords||Sevoflurane post-processing cerebral ischemia and reperfusion Bcl-2 Caspase-3|
Aim:Construct the rat cerebral ischemia and reperfusion model. Sevoflurane post-processing was used to interfere with it. To investigate the effect of sevoflurane post-processing on cerebral ischemia reperfusion as well as Bcl-2and caspase-3expression in hippocampus.Method:Fourty healthy and clean male SD rats, weighted200-250g, were provided by experimental animal center of Shanxi Medical University. They were randomly divided into four groups(10in each group). Group C:sham operation group. IP group:ischemia-reperfusion group. S1group:1.0MAC sevoflurane post-processing group. S2:1.5MAC sevoflurane post-processing group. Rat ischemia and reperfusion model was constructed by clipping bilateral carotid artery. Ischemia time in IP, S1and S2group was20minutes. Then rats in S1and S2group were reperfused immediately and inhaled with different concentrations of sevoflurane for15minutes. Rats in group C only separate bilateral common carotid arteries without clamping them. Suturing the neck incision. Rats were given free diet. Twenty four hours after reperfusion, rats were sacrificed and hippocampi were seperated. Thereafter, they were fixed by4%formalin for2h, then routine dehydration, transparency and finally paraffin-embedded. HE dyed observation cells form. Immunohistochemistry was used to detect the Bcl-2and Caspase-3protein expression. BL-2000Medical Image Analysis System was applied to exam the average gray value of Caspase-3, Bcl-2protein expression.Result:1、HE dyed:optical microscope group C cells, dyeing loose and nucleoli are clearly visible and cell membrane and nuclear membrane are complete, was pink cytoplasm. IP group of cells atrophy decrescent, intracellular dyeing is violet, inside the cell structure and the nucleus structure are disappear, intercellular increased. S1group and the S2group cellular structure exists, body of the slightly narrowed, cells in dyeing uneven crumb, visible nucleoli, normal cells mingled with lesions cells, and S1group lesions cells more.2、Caspase-3and Bcl-2protein mainly expressed in neurons. Protein immunoreactive cells showed brown cytoplasmic coloration. Bcl-2showed a uniform diffused staining.3、Bcl-2expression:The average gray value in IP group was significantly higher than other groups, which indicated a significantly reduced expression (P<0.05). The average gray value in S1and S2group was lower than other groups, which suggested a significantly increased expression (P<0.05). The average gray value of S2group was lower than that of S1group, representing the increased expression (P<0.05).4^Caspase-3expression:The average gray value of the IP group was significantly lower then other groups, which indicated a significantly increased expression (P<0.05). The average gray value of S1and S2group was significantly elevated, which suggested a reduced expression (P<0.05). The average gray value of S2group was higher than that of S1group, representing the decreased expression (P<0.05).Conclusion:1. Post-processing of sevoflurane has a protective effect on cerebral ischemia reperfusion injury.2. The mechanism was related to increased Bcl-2expression as well as reduced caspase-3expression.3. The protective effect is in dose-dependent manner.