| Objective: To establish the models of rats with intracranial hemorrhage(ICH),and given ganglioside intervention,by observing the nerve function score?the different period of brain water content(BWC)?morphology surrounding hematoma tissue,the expression of Nogo A and Ng R in the brain of rats within different times,to discuss the influence of ganglioside on the expression of Nogo A and Ng R after intracerebral hemorrhage and the possible protection mechanism of ganglioside.Methods: 1.The method of autologous blood injection to establish the rat caudate nucleus hemorrhage model.2.75 male SD rats were randomly divided into 3 groups: Sham group(25),cerebral hemorrhage group(ICH group)(25),ganglioside intervention group(GM group)(25),each group according to the execution time divided into 1d,3d,7d,14 d,28d five subgroups,there are 5 rats in each subgroup,Sham group rats only pin injection without autologous blood.3.Drug intervention: Rats in the GM group were injected intraperitoneally with30mg/kg of ganglioside immediate after operation,and then at the same time every day by the same dose of medication until death,Sham group and ICH group rats were given the same amount of saline solution at the same time by intraperitonral injection.4.Two hours after modeling,we would evaluate whether the model was successful by Bederson scoring method and evaluate nerve function score at each time point by Garcia scoring method.Water contents of a part brain tissue arounding the hematoma were observed at different time points by wet-dry-weight method,the other of rat brain was saved in 4% neutral Formaldehyde Solution,for paraffin embedding,slicing,HE staining observation of brain tissue morphology changes around hematoma,the expression of Nogo-A and Ng R protein in the brain tissue around the hematoma was detected by immunohistochemical method.Result: 1.The behavioral score: No obvious neurological dysfunction was observed in Sham group;The neurological deficits score of GM group and ICH group is lower compared with Sham group at each time point;the score of GM group contrast to ICH group is higher at 1d,3d,7d,14 d,21d group;Garcia's score was lowest at 3d group whether in ICH group or GM group,had statistics obviously(P<0.05).2.BWC: The water content of brain tissue arounding brain hematoma of ICH group and GM group were much more than the Sham group;the 3d subgroup in ICH group reached the peak;The water content showed lower level in GM group than ICH group at 1d?3d,7d,14 d,21d subgroup,had statistic obviously(p<0.05).3.Brain tissue histomorphological changes: Under the light microscope,the structure of the brain tissue of Sham group was completet,the arrangement of neurons was neat and the cell morphology was normal,no obvious nerve cell swelling and inflammatory cell infiltration;At different time points after cerebral hemorrhage,ICH group and GM group can be seen in varying degrees of brain tissue congestion and edema,part of the nerve cell swelling and was vacuolar degeneration,the nucleus was dissolved and solid shrinkage,glial cell proliferation and inflammatory cells infiltration can be seen in the stroma,the chromatin density increases,eosin staining deep.Inflammatory response and pathological changes reached the peak at 3d,and then gradually decreased;Compared with ICH group,the inflammatory response and pathological changes were lighten in the GM group at each time point.4.Immunohistochemistry showed: The sham operation group had only a small amount of Nogo A expression at each time point after operation,there was no significant difference in the expression of Nogo-A between the remaining two groups at 1d after operation(p>0.05),the 3d,7d,14 d and 21 d were significantly higher than those in the Sham group;In ICH group and GM group,the expression of Nogo A were increased at 1d,reached the peak at 7d,then decreased gradually,and there was still many expression at 21d;Compared with ICH group,the expression of Nogo-A in GM group was not significantly different at 1d after cerebral hemorrhage(p>0.05),but at 3d,7d,14 d and 21 d were obviously reduced,the above results were statistically significant(P<0.05).The sham operation group had only a small amount of Ng R expression at each time point after operation,there was no significant difference in the expression of Ng R between the remaining two groups at 1d after operation(p>0.05),the 3d,7d,14 d and 21 d were significantly higher than those in the Sham group;In ICH group and GM group,the expression of Ng R were increased at 1d,reached the peak at 14 d,then decreased gradually,and there was still many expression at 21d;Compared with ICH group,the expression of Ng R in GM group was not significantly different at 1d after cerebral hemorrhage(p>0.05),the rest of the time points were all obviously reduced,the differences were statistically significant(P<0.05).Conclusions: 1.Ganglioside can effectively reduce the degree of inflammatory response and pathological changes after cerebral hemorrhage,reduce brain edema,and promote the recovery of neurological function in rats.2.Ganglioside can effectively reduce the expression of Nogo-A and Ng R in brain tissue around the lesions after cerebral hemorrhage,thus promoting nerve regeneration and finally reaching neuroprotective effect. |
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