| Aim:To research the effect of dl-3n-butylphthalide (NBP) on the neuroprotective effect and expression of caspase-8 in the rat model of focal cerebral ischemia-reperfusion.Methods:180 male wistar rats were randomly divided into sham operation group (n=36),ischemia-reperfusion(I/R) group(n=36), NBP treatment group (n=36), NBP prevention group (n=36) and NBP prevention and treatment group(n=36). Each respectively in 1 h after ischemia-reperfusion 6h,12h and 24h for detection. Each point have 12 rats. The rat model of focal brain ischemia-reperfusion was established with the suture-occluded method introduced by improvement Zea Longa. Baseed on single-blind method Zas Longa scoer and TTC staining to determine the success of modeling modeling, with HE staining brain tissue observation pathological changes, TUNEL detection of neuronal apoptosis, Resistance observed by immunohistochemistry the expression of caspase-8 protein, Western blot and quantitative detection of each group the level of caspase-8 protein expression.Results:1. Sham-operated rats without any neurological deficiency symptoms; Ischemia reperfusion group rats with varying degrees of neurological deficit symptoms. Observation indicates, NBP treatment group, prevention group and prevention and treatment group are able to prevent improvement of varying degrees of cerebral ischemia-reperfusion injury of the symptoms of neurological deficits.2. HE staining showed the morphology of neurons of Sham group was normal. In I/R group, the ischemic side of the organizational structure of disappeared or markedly abnormal, Reduction in the number, cell disarrangement. NBP treatment group, prevention group, prevention and treatment group to varying degrees, to prevent a reduction in brain tissue necrosis. Under the light microscope, observed prevention group results slightly worse.3. Apoptotic cell showed nuclear in the TUNEL assay. There were few positive cells in Sham group, while in I/R group can see that a large number of apoptotic cells (P<0.05). Compared with the I/R group, NBP treatment group, prevention and treatment groups could inhibit neuronal apoptosis. Prevention group can also inhibit neuronal apoptosis, but the result was slightly worse than the previous two groups.4. There was occasionally interspersed expression of Caspase-8 protein in Sham group. Ischemia-reperfusion group at 6h after modeling a large number of Caspase-8-positive cells, Extended gradually increased over time (P<0.01), NBP treatment group, prevention group and preventive treatment group compared with Caspase-8 positive cells were significantly reduced than ischemia-reperfusion group (P<0.01). NBP preventive treatment group compare with the NBP treatment group, In the ischemia-reperfusion 6h,12h, expression of caspase-8, the treatment group loose than preventive treatment group (P<0.05), Between the two groups at 24 h, expression of this protein was no significant difference (P> 0.05).5. Western blot:Normal rats and sham-operated group, Caspase-8 activity in brain tissue fragments of protein expression is very low. 1h ischemia-reperfusion 24h, the ischemia-reperfusion group the expression of Caspase-8 protein was significantly raised (P <0.01). NBP treatment group, prevention group and the prevention and treatment of the three groups of this protein expression was significantly lower than that ischemia-reperfusion group (P<0.01), but the NBP treatment group decreased expression of Caspase-8 protein, more pronounced (P<0.05); Prevention and treatment groups in the ischemia-reperfusion 24h of this protein expression and no significant difference between treatment groups (P> 0.05).Conclusions:1. NBP prevention and treatment could attenuate pathological change after cerebral ischemia reperfusion, show a neuroprotective effect.2. NBP obviously inhibited transient focal cerebral ischemia induced apoptosis in ischemic penumbra area, relieve neuronal injury.3. NBP can decrease neuronal apoptosis, inhibit the expression Caspase-8 protein in the cerebral ischemia and reperfusions, so as to protect the brain. |
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