The Expression Of Cytochrome-c And Its Significance In The Treatment Of Butylphthalide On Cerebral Ischemia-reperfusion Injury

Objective:Cerebral ischemia/reperfusion injury and programmed cell death have close relation,mitochondria constitute the control center of survival, in the mitochondria-dependent apoptosis pathway,cytochrome-c released from mitochondria into the cypoplasm is the important process of apoptosis.Butylphthalide is the first chemical drug with independent intellectual property rights in China.In this study, using different methods to observe the expression of cytochrome-c in Butylphthalide on focal cerebral ischemia-reperfusion injury,and explore Butylphthalide protective effect on nerve cells, provide experimental evidence for prevention and treatment of cerebrovascular disease.Methods: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+treatment group(n=36),each group is further divided into ischemia-reperfusion 6h,12h,24h three sub-groups.Using modified method to make focal cerebral ischemia-reperfusion model. 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+ treatment group;in sham-operation group and model group, inject the same saline after 1h ischemia. Brain pathology with HE stainting,observe the change of cytochrome-c in brain tissue with immune staining,detect apoptotic cells with TUNEL.Results:1.Using modified suture method can produce MACO/R model successfully,focal ischemic brain tissue can be observed obviously.2. There are different degrees of edema and cavity formation scattered in the ischemic brain tissue of cerebral ischemia-reperfusion model group,the number of apoptotic cells increased significantly. There are varying degrees of improvement in prevention and treatment group.3.Compare with sham-operated group,cytochrome-c expression and the number of apoptotic cells increased significantly in the cerebral ischemia-reperfusion model group;in the prevention group, treatment group, prevention+treatment group,cytochrome-c and apoptotic cells were decreased to varying degrees, the effect from high to low is the prevention+treatment group,the treatment group,the prevention group.Conclusions:1.Focal cerebral ischemia-reperfusion model was produced by suture method is reliable and effective.2.Brain tissue pathological changes severly,cytochrome-c expression and apoptotic cells increased after cerebral ischemia-reperfusion.Infer that the large cytochrome-c expression may be one of the important mechanisms about cerebral ischemia-reperfusion injury.3.Butylphthalide on focal cerebral ischemia-reperfusion injury has obvious protective effects, including prevention and treatment.4.Butylphthalide may play its neuroprotective effect by inhibiting the release of cytochrome-c.