| Objective:Through the establishment of rat cerebral hemorrhage(ICH)model,to investigate the effect of nerve growth factor(NGF)on the expression of IL-6 and SOCS-3 after intracerebral hemorrhage in rats,and possible protective mechanism.Methods:1 ?Establishment of intracerebral hemorrhage model in the caudate nucleus of rat by non heparin autologous blood injection.2?80 healthy male SD rats were randomly divided into four groups:Sham group(5),cerebral hemorrhage group(ICH group)(25),small dose of nerve growth factor in the treatment group [NGF(S)group](25),large dose of nerve growth factor in the treatment group [NGF(L)group](25).The sham operation group rats were inserted pin without blood and time point only set as 6h after surgery,the ICH group,NGF(S)and NGF(L)group were divided into 5 subgroups according to the time points: 6h,24 h,2d,3d and 5d,each of the 5 rats in each subgroup.3?Method of administration:The rat model of intracerebral hemorrhage was established 30 minutes later,the NGF size dose group,NGF(S)group were given 3ug/kg,NGF(L)group each time to give 18ug/kg.For the first time double with routine dose of intraperitoneal injection,after every day at the same time according to the routine dose.Rats in group sham and group ICH were injected with the same amount of normal saline at the same time.4?The evaluation of 2h after operation was evaluated,the neurological function scores were evaluated at each time point after operation,determination of water content in brain tissue around hematoma by wet and dry weight method,HE staining method was used to observe the inflammatory reaction and pathological changes of the brain tissue around the hematoma,immunohistochemistry were used to observe the expression of IL-6 and SOCS-3 positive cells in the brain tissues around the hematoma,TUNEL staining were used to observe the apoptosis of the brain tissue around the hematoma.Results:1?Behavioral score: there was no obvious neurological dysfunction in Sham group rats;The neurological function scores of ICH group and NGF group were lower than Sham group at different time points after cerebral hemorrhage operation(P<0.05);ICH group?NGF(S)group and NGF(L)group in the cerebral hemorrhage after operation 24 h,6h,5d each time point and 2d time point comparison difference has statistical significance(P<0.05);NGF(S)group,24 h,2d,3d,5d time points after cerebral hemorrhage were compared with the ICH group,the difference was significant(P<0.05).The neurological function scores of NGF(L)group at each time point were higher than that in group NGF(S)and ICH group(P<0.05),Which compared with the cerebral hemorrhage group,the difference was more obvious(P<0.05).2?Water content of brain tissue:ICH group and NGF group rat cerebral hemorrhage cerebral tissue around the hematoma water content at each time point were obvious higher than sham group(P<0.05);ICH group?NGF group after cerebral hemorrhage 24 h,6h,5d each time point and 2d comparison has statistical difference(P<0.05);NGF(S)group at 24 h,2d,3d,5d each time point of brain tissue water content was lower than the ICH group(P<0.05);The brain tissue water content in NGF(L)group was lower than that in NGF(S)group and ICH group(P<0.05),Which compared with the cerebral hemorrhage group,the difference was more remarkable(P<0.05).3?Inflammatory reaction and pathological changes:Sham group showed that the structure of the brain tissue was clear and complete,and the morphology and structure of the cells were in good condition,neurons arranged neatly,no obvious inflammatory cell infiltration and cell swelling;ICH group and NGF group at each time point after cerebral hemorrhage showed different degree of brain tissue congestion and edema,nerve cell swelling.In the form of vacuole degeneration,degeneration and necrosis of the nucleus,dissolution of the nucleus,and the widening of the gap between the cell and the blood vessel,inflammatory cell infiltration and glial cell proliferation,increase in the density of the chromatin,deep red colorin.2-3d inflammatory reaction and pathological changes to reach the peak,then gradually reduced;Compared with group NGF,the degree of inflammatory reaction and pathological changes of ICH group were decreased at the corresponding time points,the NGF(L)group was more significant for the improvement of inflammatory reaction and pathological changes.4?Expression changes of IL-6 positive cells:in group Sham,only a few IL-6 positive cells were expressed in the rats after operation;ICH group and NGF group of rats with cerebral hemorrhage at each time point have different degrees of positive cell expression,compared with the Sham group,the difference was significant(P<0.05);NGF and ICH group in cerebral hemorrhage after 6h,24 h,3d,5d each time point,the expression of positive cells of IL-6 compared with 2d,the difference was significant(P<0.05);The expression of NGF(S)group at each time point in IL-6 cells after cerebral hemorrhage operation is lower than that of group ICH(P<0.05);The expression of IL-6 positive cells in NGF(L)group was lower than that in ICH group and NGF(S)group(P<0.05).5 ? Expression changes of SOCS-3 positive cells:In sham group,only a few SOCS-3 positive cells were expressed in the rats after operation;ICH group and NGF group of rats with cerebral hemorrhage at each time point have different degrees of positive cell expression,compared with the Sham group,the expression of SOCS-3 positive cells was significantly increased(P<0.05);In ICH group and NGF group,there were statistically significant differences in 6h,24 h,5d positive cells at each time point of SOCS-3 after intracerebral hemorrhage compared with 2d(P<0.05),;NGF(S)in the cerebral hemorrhage group at each time point after operation the number of positive cells expressing SOCS-3 were increased compared with ICH group(P<0.05);The expression of SOCS-3 positive cells in NGF(L)group was higher than that in ICH group and NGF(S)group(P<0.05);Compared with the cerebral hemorrhage group,the difference was more significant(P<0.05).6?Comparison of apoptosis of brain tissue around hematoma:TUNEL staining showed no obvious apoptosis cell expression in Sham group;ICH group and NGF group had different degrees of apoptosis after intracerebral hemorrhage,ICH group,NGF group and Sham group at the corresponding time points after cerebral hemorrhage apoptosis,the difference was significant(P<0.05);The expression of apoptosis cells in ICH group and NGF group was statistically significant at each time point of 6h,24 h,3d,5d after operation compared with 2d(P<0.05);NGF(S)group and ICH group at each corresponding time point cell apoptosis was reduced(P<0.05);NGF(L)group and ICH group,NGF(S)at the corresponding time,the number of apoptotic cells were lower(P<0.05),compared with the cerebral hemorrhage group,the difference was more obvious.Conclusion:The rat tail autologous blood injection method to establish rat model of intracerebral hemorrhage,the pathophysiological changes and lead to human body after cerebral hemorrhage is consistent,it is an ideal animal model of cerebral hemorrhage.2?After cerebral hemorrhage occurred in brain edema and brain tissue inflammation and apoptosis,to aggravate neurological dysfunction.3 ? After cerebral hemorrhage,IL-6 SOCS-3 expression increased,NGF can increase the expression of SOCS-3 is more obvious,at the same time,the expression of IL-6 can be reduced.4?NGF may can up regulation of SOCS-3 expression and down regulated IL-6 expression,reduce cerebral edema,and cell apoptosis,it has protective effect on nerve function,and the effect of large dose of NGF is better than that of small dose. |
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