The Effect Of Motor Training Beginning At Different Time To The Expression Of BDNF Protein, IL-10 Protein, Bax Gene And Bcl-2 Gene In The Motorcortex Around The Hematoma After Intracerebral Hemorrhage In Rats

objective: This article trys to explore the possible mechanisms of motor therapy which improves the motor function of rats after intracerebral hemorrhage and the effect of intervening motor training in different time to the functional plasticity and apoptosis of rats after intracerebral hemorrhage.It could provide guidance about the intervention time of motor training for intracerebral hemorrhage patients in clinical.Methods : 1 ? The methods of molding and intervention: we made the model of intracerebral hemorrhage by injecting the collagenase of ? type and heparin sodium to the caudate nucleus of rats.On the contrary,we injected the same volume of normal saline into the caudate nucleus of rats in the control group(sham-operated group).We adopted Modified Neurological Severity Scores(mNSS)and micro PET/CT to evaluate the models.The rats were divided into 24 hours(24 h)group,3 day(3d)group and 7 day(7 d)group depending on the different starting time of motor training after the success of molding.Each group was trained for 3 weeks.The control group and model group were not given any treatments.2 ?observation index:(1)The changes of the motor function in rats after the intervention were evaluated by the test of rolling-bar,screen and balance beam;(2)We observed the changes of histopathological of cerebral tissue around thehematoma by HE staining,and detected the expression of brain derived neurotrophic factor(BDNF)and interleukin 10(IL-10)in motor cortex around the hematoma by immunohistochemical staining;(3)We detected the expression of Bcl-2 mRNA and Bax mRNA in motor cortex around the hematoma by RT-PCR.3?Statistical method: Statistical analysis was performed with IBM SPSS Statistics 17.0 software and all the data was indicated in the form of Mean±standard deviation(?±s).One-way analysis of variance(one-way ANOVA)and SLD-t test were used to compare the average optical density value of IL-10 protein and BDNF protein between all the groups.The same as to the Grey value of Bax mRNA and Bcl-2 mRNA and the ratio of Bcl-2 mRNA and Bax mRNA.p<0.05 has statistical significance and p<0.01 with significant statistical meaning.The correlation test was undertaken with the linear correlation analysis.Results: 1?motor function score: The scores of the motor training groups were all lower than the model group in the three tests(p<0.05).But there was no significant difference between the three motor training groups.The three tests of motor function in the control group were zero.2?HE staining: The cells in the right caudate nucleus of rats in control group were normal.The hematoma in model group has basically disappeared and left a cavity.The cerebral tissue around the cavity was loose and we could see many vacuoles of different sizes in it and accompanied by the glial cells proliferation and angiogenesis.The angiogenesis around the hematoma in 24 h group was performed more obviously than the model group.The angiogenesis around the hematoma in 3d group and 7d group performed more obviously than 24 h group,but the degree of loose in cerebral tissue around the cavity was improved and the number of vacuoles was decreased.3 ? The expression of BDNF: The expressions of BDNF in any of the three motor training groups were higher than the model group(p<0.01).In the 3d group the expression of BDNF was the highest,the second was 24 h group and the last was 7d group,the differences between the three motor training groups had significant statistical meaning(p<0.01).4?The expression of IL-10: The expression increased in the motor training groups.Any of the motor training groups has significant statistical meaning compared with model group(p<0.01).In the three motor training groups,the expression in the 3d group was the highest and the differences compared with the 24 h group and 7d group were significant statistical meaning(p<0.01).However,there was no significant difference between 24 h group and 7d group(p>0.05).5?The expression of Bax mRNA and Bcl-2 mRNA:(1)Bax mRNA: The expression of Bax mRNA in the three motor training groups were all higher than that in model group(p<0.01).The expression in 7d group was higher than that in the 24 h group and the 3d group(p<0.05).However,the difference between the 24 h and the 3d group had no statistical significance(p>0.05).(2)Bcl-2 mRNA: There was no obviously increased in the expression of 24 h group compared with model group(p>0.05).The expression in the 3d group and the 7d group were higher than the model group(p<0.05).However,the differences among the three motor training groups were small(p>0.05).(3)Bcl-2 mRNA/Bax mRNA: The ratio of Bcl-2mRNA and Bax mRNA in motor training groups were higher than that in modelgroup(p<0.01).The ratios in 24 h group?3d group and 7d group were increasing gradually.The ratio of 7d group risen in the most obviously and compared with24 h group and 3d group had statistical significance(p<0.05).However,the difference between the 24 h and 3d group had no significance(p>0.05).6 ?Correlation analysis: The expression of IL-10 protein and BDNF protein were positively correlated and the expression of IL-10 protein and the Bcl-2 mRNA were positively correlated.Conclusion: 1 ? Taking the collagenase induced method which injecting the collagenase type I and heparin sodium to the caudate nucleus of rats can successfully make the animal models of intracerebral hemorrhage.2 ? Motor training can reduce the score of motor function and promote the expression of IL-10 protein and BDNF protein in the motor cortex around hematoma.3?Motor training can effectively improve the motor function of rats with intracerebral hemorrhage and raise the expression of Bcl-2 gene and reduce the expression of Bax gene.4? Beginning the motor training at 3 days after rats cerebral hemorrhage is conducive to the expression of BDNF protein and IL-10 protein in the motorcortex around the hematoma than 24 hours and 7 days.5?Beginning the motor training at 7 days after rats cerebral hemorrhage is detrimental to the expression of Bax gene in the motorcortex around the hematoma than 24 hours and 3 days,but not conducive to the expression of Bcl-2 gene.