| Objective:Using different methods to investigate the dynam changes of the apoptotic protein Smac and the histologic outcome of the apoptosis neuron in the rat models of focal cerebral ischemia-reperfusion injury at different time points and the impact of the NBP to the changes.Methods:1. Focal cerebral ischemia-reperfusion model was established by the occlusion of right middle cerebral artery.2.180 male Wistar rats were randomly divided into sham-operated group (n=36), model group (n=36), Butylphthalide treatment group (n=36), Butylphthalide prevention group (n=36), Butylphthalide prevention plus treatment group (n=36), each group is further divided into ischemia-reperfusion 6h,12h,24h three sub-groups. Butylphthalide measured by 80mg/kg dose. Inject Butylphthalide dilution by intraperitoneal after 1h ischemia in Butylphthalide treatment group; one week before the operation, with a continuous gavage once a day in Butylphthalide prevention group; with a continuous gavage once a day one week before the operation and inject Butylphthalide dilution by intraperitoneal after 1h ischemia in Butylphthalide prevention plus treatment group; in sham-operation group and model group, inject the same saline after 1h ischemia.3. Brain pathology with HE stainting, observe the change of Smac protein in brain tissue with immune staining, detect apoptotic cells with TUNEL.Results:1. HE staining shows that infarction and neuronal injury was relieved at different time points in the Butylphthalide treatment group and Butylphthalide prevention plus treatment group compared with model group.2. Immunohistochemical results show that: there were elevated expression of Smac protein in model group compared with sham operated group, and as ischemia-reperfusion time Smac protein expression gradually increased, at 24h after reperfusion reaching the top; the expression of Smac protein were significantly decreased in Butylphthalide treatment group and prevention plus treatment group, there were great significance in Butylphthalide treatment group and prevention plus treatment group compared with model group (P＜0.05).3. westernblot results show that:at 24h after ischemia-reperfusion the expression of Smac protein rows from high to low:model group, Butylphthalide prevention group, Butylphthalide treatment group, Butylphthalide prevention plus treatment group and sham-operated group, among them there were great significance in Butylphthalide treatment group and prevention plus treatment group compared with model group(P<0.05).4. TUNEL staining shows that there were a lot of apoptotic cells in the model group compared with corresponding time points, it was relieved greatly in the Butylphthalide treatment group and Butylphthalide prevention plus treatment group(P<0.05).Conclusions:1.The rats development typical symptoms of neurological impairment, cerebral HE staining results in line with pathological process.2. The expression of Smac protein changed dynamically, it increased in 6h, as ischemia-reperfusion time Smac protein expression gradually increased, at 24h after reperfusion reaching the top, so Smac protein may involve in the pathological process of ischemia-reperfusion, Butylphthalide could reduce the apoptosis neuron through dropping the expression of Smac protein, thereby it could protect the neuron. |
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