| Objective: In order to investigate whether the butylphthalide injectionpretreatment can go through the signaling pathway of PI3K／AKt to see theneuroprotective effect and its possible mechanism of cerebral ischemia reperfusionlesion,the author will use the method of observing butylphthalide injection pretreatment,and take the damage model of SD, MCAO, and HE staining to observehistopathological form rats, immunohistochemical method to observe the expressionchange of caspase-3and p-AKt.Methods: The author will use36healthy adult male SD rats whose weight are200-250g and randomly divide them into3groups: sham-operative group, cerebralischemia reperfusion model group (model group), cerebral ischemia reperfusion groupof butylphthalide injection pretreatment (referred to as butylphthalide injectionpretreatment group). Every team will get12rats. To take the method of drugpretreatment, the rats in the group of butylphthalide injection pretreatment will be givenbutylphthalide injection200mg/kg/d; the other two groups will be given volumetric0.9%sodium chloride normal saline, intraperitoneal injection, twice a day, for5consecutive days.The rats after pretreatment given, Zea-Longa Suture-occluded method to make ratstransient (MCAO) model.Implementing ischemia for2hours and reperfusion for24hours. The standards for successful model: after the rats waking up, they will show thephenomenon,that is, on the right side of Homer and the contralateral forelimb primarilyparalysis and neural function grade:1-3.The sham-operative group just show CCA,ECAand ICA. The rest will not do any processing. After the success of making rats molding, refer to Zea-Longa5points as thestandard to grade european stroke score. And then, randomly take6rats from everyteam, take out their brain tissue:（1）2%chlorination2,3,5-Triphenyl tetrazaliumchloride, TTC to observe the Percentage of volume of cerebral infarction;（2）Hematoxylin-eosin, HE dyeing to observe the changes of brain organization neuron;（3）caspase-3、p-AKt cell immunohistochemical staining method to calculate positivecells expression rate.Results:1. The scores of rats’ neurologic impairment: The sham-operative group does notobserve neurological impairment (0score); compared to the sham operation group, theother two groups show the obvious neurological impairmen（P<0.01）；compare to themodel groupp,the degree of neurological impairment the butylphthalide injectionpretreatment group is relatively reduce（P<0.05）.2. To determinate the rats’ volume of cerebral infarction: The brain tissues in thesham operation group after being TTC dyeing, they all show red color, and no whiteinfarcts formation; compared to this group, the other two groups have evident whiteinfarcts to form（P<0.01）；compare to the model group, the volume of white infarctsdecreased in the butylphthalide injection pretreatment group（P<0.05）.3. The rats’brain tissues HE staining:The nerve cells in the sham operation grouphave normal morphology, structural integrity, well arranged, the intercellular substancedensed without edema; the normal nerve cells in model group evidently decreased, alarge of degeneration necrosis cells, cellular structure not clear, disordered arrangement,pyknosis and deep stained nuclear, cell cytoplasm vacuoles formed, cells interstitialedema; compared to the model group,most of nerve cells in the butylphthalide injectionpretreatment group have structural integrity, the changes of cell formation are lighterand nuclear pyknosis are less, tissues edema are lighter.4. Compared to the the sham operation group, caspase-3、p-AKt positive cellsexpression are evidently increased in the other two groups（P<0.01); compare to themodel group,the caspase-3positive cells expression are decreased in the butylphthalideinjection pretreatment group（P<0.05）, but the p-AKt positive cells expression inbutylphthalide injection pretreatment group is relatively high.（P<0.05）.Conclusions:1.Butylphthalide injection pretreatment can reduce cerebral ischemia/reperfusioninjury, reduce the neural function defect scale, narrow focal cerebral infarction volume, relieve neuron pathology morphological injury.2.The method of Butylphthalide injection preconditioning can reduced the numberof caspase-3expression, increased the number of p-AKt expression, and can inhibit theapoptosis caused by cerebral ischemia/reperfusion.3.Butylphthalide injection pretreatment have nerve protective effect on cerebralischemia reperfusion injury.Mechanisms can be relevant With the raising of PI3K/AKtpathway and inhibition of apoptosis. |
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