Effect Of Dl-3n-butylphthalide On The Expression Of Caspase-9 And Neuron Apoptosis In The Rat Model Of Focal Cerebral Ischemia-referfusion

Objectives: To investigate the effect of dl-3n-butylphthalide(NBP) on the histologic outcome,apoptosis and expression of apoptosis regulating protein caspase-9 at different time points on the model of focal cerebral ischemia-reperfusion injury in rats.Methods:1. Focal cerebral ischemia-reperfusion model was established by the occlusion of right middle cerebral artery . 2. Healthy male Wistar rats were randomly divided into sham group, ischemia–reperfusion( I-R)group, NBP treated group, each 18. Each respectively in 1 h after ischemia-reperfusion 6 h, 12 h and 24h for detection. HE staining of pathological changes in brain tissue morphology; immunohistochemical methods to detect the Caspase-9 protein expression changes; TUNEL staining of the group to observe the neuronal apoptosis.Results: 1.HE staining shows that infarction and neuronal injury was relieved at different time points in the NBP treated group compared with I-R group. 2. Immunohistochemical results show that:there were elevated expression of caspase-9 in I-R group compared with sham operation group, and as ischemia-reperfusion time Caspase-9 expression gradually increased,at 24h after referfusion reaching the top. expression of Caspase-9 in NBP treated group at all time points were significantly decreased(P <0.01). 3. A large number of TUNEL-positive cells were observed in I-R group, the number of apoptotic cells was higher than that of sham-operated group (P<0.01). TUNEL-positive cells of cerebral cortex and striatum were markly reduced in NBP treated group (P<0.01).Conclusions:1. The rats development typical symptoms of neurological impairment.TTC staining showed infarction.Cerebral HE staining results in line with pathological process. 2.NBP could attenuate cell death after cerebral ischemia-referfusion, show a neuroprotictive effect. 3. NBP obviously inhibited transient focal cerebral ischemia induced apoptosis in cortex and striatum, decrease the activation of caspase-9 , block activation of caspase cascade and relieve neuronal injury.