Research On Influence Of The Expression Of Cell Apoptosis Related Proteins In Experimental Intracerebral Hemorrhage In Rat With Ganglioside Intervention

objective: We establish the cerebral hemorrhage model,use ganglioside intervention, detect the expression changes of apoptosisregulatory proteins within the tissue around hematoma after intracerebralhemorrhage: the proteins are Fas、FasL、Bax、Bcl-2. Discuss gangliosideprotective effect on brain cells. Methods:1.grouping methods:45maleSD rats weight only for250~300grams, It is divided into3groups atrandom: the control group (n=8), intracerebral hemorrhage group(ICHgroup)(n=20), intracerebral hemorrhage and ganglioside interventiongroup(GM1group)(n=20), each group according to the execution timedivided into6h,24h,3d,7d four subgroups.2. Preparation of hemorrhagerat model: Use brain stereotaxic instrument preparation method ofautologous blood injection hemorrhage rats model. Cut the tails of therats of ICH group and GM1group and get the blood, inject the blood intothe brain of the rats to making model of cerebral hemorrhage. The controlgroup steps are the same as the other two groups but inject into the blood.3. Model successful evaluates.4. Drug intervention: Inject ganglioside30mg/kg into rat abdominal cavity in GM1group immediate after operation.ICH group and control group with the same amount of normal salineinstead.5. Evaluation of nerve dysfunction: Garcia grading method is adopted to improve the score,3point is minimum,18point is the highest.The higher scores show the lighter neural dysfunction, the lower the score,the heavier neural function damage.6. Specimen extraction and detection:all groups are at specified time points to be put to death. Cut off the headand get the brain, make the brain tissue slices. Use TUNEL stainingmethod to determine the apoptotic cells of the brain tissue. Detect theapoptosis related proteins of Bax/Bcl-2/Fas/FasL withimmunohistochemical method and analyzed its average integral opticaldensity (AIOD value).7. Statistical analysis: Using analysis of varianceand linear correlation analysis. Result:1.Nerve function obstacle ofpostoperative of GM1group light than ICH group (P <0.05).2. TUNELstaining showed that apoptosis ICH group and this group significantly inthe control group (P <0.05); Cell apoptosis of GM1group are less thanthat in the ICH group (P <0.05).3. The immunohistochemical display: ineach time point the expression of apoptosis Promotion proteins as Bax,Fas, FasL of GM1group is lower than that in ICH group (P <0.05); ineach time point the expression of apoptosis Suppression protein as theBcl-2of GM1group is higher than that in ICH group (P <0.05).4.Linear correlation analysis: Bax protein expression is positivelyassociated with brain cells apoptosis (r=0.821, P <0.01); The Bcl-2protein expression and brain cells apoptosis shows a negative correlation(r=0.666, P <0.01); Fas protein expression is positively associated with brain cells apoptosis (r=0.415, P <0.01); FasL protein expression ispositively associated with brain cells apoptosis (r=0.775, P <0.01).Conclusion:1. The apoptosis mechanism involves in the secondary braininjury after intracerebral hemorrhage.2. Ganglioside has significantlynerve protective effect on intracerebral hemorrhage rat, its mechanismmay be that: decrease the expression of the proteins which can promotecell apoptosis, increase the expression of the proteins which can restraincell apoptosis.