| Objective: To observe the variation of expression of GFAP in the lesion areas after acute injury of spinal cord and to analyze the regulation of GFAP on endogenous neural stem cells to ensure the molecule treatment mechanism of Ligustrazine for spinal cord.Methods: The SD(Sprague Dawley)rats were randomly divided into gap group(group A),pseudo surgery group(group B),model group(group C),and Ligustrazine treatment group(group D).The rat models of acute spinal cord injury were established by modified Allen's method in the experiment.The rats of group D after molding were given abdominal Ligustrazine Hydrochloride Injection for 200 mg/kg per day,and the first injection was given after 30 min after the operation.The group A,B and C were given the same amount of physiological saline in abdomen at the same time for ten days while group D was given the same mount with the former three groups in the following nine days after molding on the second day.Oblique board test and BBB evaluation on nerve function were applied to test the postoperative results on the 8th hour,1st day,3rd day,7th day,14 th day,and 28 th day respectively.6 rats in each group were sacrificed on the 8th hour,1st day,3rd day,7th day,14 th day,and 28 th day after spinal cord injury.Then the injuried spinal cord about the 5mm right and left of it was cut out to be detected for the Nissl staining and immunohistochemistry.Meanwhile,the experiment adapted Nissl staining methods to observe the morphological changes of spinal cord.Immunohistochemistry was applied totest the expression of Brdu+ and Nestin+ cells and GFAP in the spinal cord.All of the test results were statistically analysis,and the correlation analysis were made among the expression of Brdu+ and Nestin+ cells,GFAP in the spinal cord.Results:Firstly,motor function of hind limbs and the function of spinal cord of the model control group and the Ligustrazine treatment group rats decreased significantly after spinal cord injury;The maximum angles the rats maintained on oblique board in the Ligustrazine treatment group were on the uprising trend and there was significant difference compared with model control group on the 14 th day and 28 th day(P< 0.01).There was significant difference compared with model control group on the 3th day,14 th day and28 th day according to the BBB score(P<0.01).Secondly,the experiment adapted Nissl staining methods to observe the pathological changes of spinal cord.Visible damage to the spinal cord edema after spinal cord injury,such as focal hemorrhages,neuron cell shrinkage,nuclear pyknosis,nissl body began to decline and disappear,neuron degeneration,3 days later,spinal cord edema,focal hemorrhages scope and inflammatory cells infiltration increased,gray matter tissue dissolved and formed cavity;To the 7th day,14 th day,necrosis range further expand,inflammatory edema gradually reduce,to the 28 th day,the scar hyperplasia was around the necrotic area and then fill in it.Ligustrazine could remarkably reduce edema,inflammatory cell infiltration and blooding and promote the repair of spinal cord tissues after injury.Thirdly,Brdu+ cells express in Gray and White Matter and the central ventricular zone,the expression of Brdu+ cells was increased when they were activated after spinal cord injury.there was significant difference(P<0.01)on the 7th day,14 th day and 28 th day after spinal cord injury sinceLigustrazine enabled to increase the proliferation of Brdu+ cells(compared with group C P<0.01).Ligustrazine could conspicuously enhance the expression of Nestin+ cells and reach its summit on the 7th day and there was significant differencecompared with group C on the 7th day,14 th day and 28 th day after spinal cord injury(P<0.01).In the experiment,the expression of GFAP increased to the most on the 7th day,and then gradually decreased,but still higher than the normal value.There is obvious difference between among Ligustrazine treatment group and model control group at each time point(P<0.01).Finally,The correlation analysis was performed on the expression of GFAP,Nestin+ cells and Brdu+ cells in the spinal cord.GFAP is positively related to Nestin+ expression.Conclusion: Firstly,the expression of GFAP positively regulates the proliferation and differentiation of endogenous neural stem cells.Secondly,Ligustrazine could promote the proliferation and differentiation of endogenous neural stem cells after spinal cord injury.Thirdly,Ligustrazine could promote the expression of GFAP. |
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