| Objective: To explore the effect of butylphthalide(NBP)on apoptosis related protein Bcl-2 and Bax and the possible mechanism of treating intracerebral hemorrhage(ICH),we observe the effect of different doses of butylphthalide on neurologic deficit,pathological change of brain,brain water content,apoptosis around hematoma tissue and the expression of Bcl-2and Bax in rats with intracerebral hemorrhage,for providing the new direction and theoretical basis for the clinical treatment of butylphthalide.Methods:200healthy adult male SD rats were randomly divided into Sham group(Sham operation group)with 50 rats,ICH group(Intracerebral hemorrhage group)with50 rats,small dose group(Small dose therapy group of butylphthalide)with 50 rats,high dose group(Large dose treatment group of butylphthalide)with 50 rats.The rat model of ICH was established by injecting autologous blood into caudate nucleus in ICH group,small dose group and high dose group.In sham group,only harnpan was drilled hole without autologous blood.The method of Berderson was used to evaluate the success of ICH model.Six hours,one day,three days,five days and seven days after modeling were the observation points and execution points,and the four groups were divided into 5subgroups with 10 rats of each group.Small dose group and large dose group were injected intraperitoneally with butylphthalide injection of 2.5mg/kg and 10mg/kg immediately after successful modeling according to body weight,twice a day,till the corresponding time point.the nerve function of the rats was scored by Garcia scoring method.After the rats were scored at each time point,the brain was quickly cut off,dry-wet method was used to measure the water content of Brain tissue around hematoma,HE was used to observe the pathological changes,TUNEL method was used to measure apoptosis and the expression of Bcl-2 and Bax protein was detected by immunohistochemistry.Finally,statistical analysis was carried out.Results:(1)Neurological deficit score: Neurological deficit score in ICH group,small dose group and high dose group was lower than that of the Sham group at each time point,and the differences were statistically significant(P<0.05).Neurological deficit score of each bleeding group became the lowest at 3d.The scores of neurological deficit in small dose group and large dose group were higher than those in ICH group at each time point,and the differences were statistically significant(P<0.05).The scores of high dose group were higher than that of small dose group except for the 6h time points,and there are statistic differences(P<0.05).(2)Water content of brain tissue: The water content of brain tissue in ICH group,small dose group and high dose group was higher than that in the Sham group except for the 7d time points,and the differences were statistically significant(P<0.05).The water content of brain tissue was the highest in each bleeding group at 3d.The water content of brain tissue in small dose group and high dose group was significantly lower than that in ICH group except for the 7d time points,there are statistic differences(P<0.05).The water content of brain tissue in the high dose group was lower than that in small dose group except for the 7d time points,and the differences were statistically significant(P <0.05).(3)Morphological changes of brain tissue: No hematoma was found in brain tissue of Sham group.No obvious tissue structure abnormality was observed under light microscope.Varying degrees of hyperemia,inflammatory cell infiltration and neuronal swelling were observed after intracerebral hemorrhage in ICH group,low dose group and high dose group.Focal neuronal cells appeared degeneration and necrosis with glial cell proliferation,and pathological changes and inflammatory reaction gradually aggravated with the peak at 3d accompanied by partial liquefaction necrosis of brain tissue.Then the pathological changes gradually decreased.the pathological changes in the small dose group and the large dose group were less than those in the ICH group after 1d,and the large dose group was more obvious.(4)Apoptosis: The number of apoptotic cells in ICH group,small dose group and high dose group was more than that in the Sham group at each time point,and the differences were statistically significant(P<0.05).The number of apoptotic cells in 3d was the most in each bleeding group.The number of apoptosis in small dose group and high dose group was lower than that in ICH group at each time point with the statistically differences(P<0.05).The high dose group was lower than the small dose group at each time point with the differences of statistically significant(P<0.05).(5)Bcl-2 protein:The amount of Bcl-2 positive protein expression in ICH group,small dose group and high dose group was more than that in the Sham group at each time point,and the differences were statistically significant(P<0.05).The expression of Bcl-2 positive protein of 3d groups was most in each hemorrhage group.The amount of Bcl-2 positive protein expression in low dose group and high dose group was higher than that in ICH group at each time point with the statistically differences(P<0.05).And the high dose group was higher than the small dose group except for the 6h time points with the differences of statistically significant(P<0.05).(6)Bax protein:The amount of Bax positive protein expression in ICH group,small dose group and high dose group was more than that in the Sham group at each time point,and the differences were statistically significant(P<0.05).The expression of Bax positive protein of 3d groups was most in each hemorrhage group.The amount of Bax positive protein expression in small dose group and high dose group was lower than that in ICH group at each time point with the statistically differences(P<0.05).And the high dose group was lower than the small dose group except for the 6h time points with the differences of statistically significant(P<0.05).Conclusions:(1)The animal model of intracerebral hemorrhage adopted autologous blood injection.The procedure was convenient and simple with high success rate,good stability,strong reproducibility,and the changes of pathophysiology and nerve function of the model were similar to those of human cerebral hemorrhage.So it is an ideal animal model of intracerebral hemorrhage and canbe widely used.(2)After intracerebral hemorrhage,the expression of Bcl-2and Bax around hematoma were obviously increased with the obviously increased number of neuronal apoptosis,and the brain edema and pathological changes of brain tissue were obvious,which resulted in neurological dysfunction.(3)Butylphthalide can up regulate the expression of Bcl-2,down regulate the expression of Bax,reduce the apoptosis of neurons after intracerebral hemorrhage,and achieve the protective effect of brain.(4)The neuroprotective effect of butylphthalide in the treatment of intracerebral hemorrhage is enhanced with the increase of dose,and the high dose butyphthalide is better than the low dose. |
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