| Objective: The spinal cord injury model of rats was treated with Balanophora polysaccharide,and the pathological changes of injured spinal cord tissue before and after treatment were observed and compared.The mechanism of protective effect of Balanophora polysaccharide on spinal cord injury was investigated by detecting the corresponding autophagy related proteins.Methods: 150 male Sprague-Dawley rats were randomly divided into sham-operation group,model group and Balanophora polysaccharide treatment group.Then,the model group and the sham operation group were given 2ml saline daily,and the Balanophora polysaccharide treatment group were given three different doses of Balanophora polysaccharide solution 25mg/kg,50mg/kg,100mg/kg respectively.Finally,the behavioral BBB score was used to evaluate the therapeutic effect of Balanophora polysaccharide on spinal cord injury.The effects of Balanophora polysaccharide on motor function of rats after spinal cord injury were evaluated by using Nissl’s staining method and comparing the histopathologic changes of spinal cord tissue of rats after1,3 7days of treatment.Immunohistochemical staining was used to detect and compare the expression of LC3 protein in the spinal cord.Western Blotting was used to detect the expression of the corresponding autophagy associated protein Beclin1,LC3-Ⅱ p62.Results: 1 BBB score: the sham operation group at different time BBB were close to the normal value;the Balanophora polysaccharide treatment group compared with model group,the high dose group after 3,7 days were higher than that of model group(p<0.05),middle dose group on the 7 day after operation was higher than in the model group(p<0.05),and,there was no significant difference between the high dose group(p>0.05).The low dose group and model group had no difference.2 Nissl staining results: sham operation group Nissl body particle number more,and evenly distributed in the cytoplasm,showing a deep purple color,a relatively good shape.Neurons compared with sham operation group,model group,each time the spinal cord neurons are not complete,and the cell gap increased,neurons also on the shrinkage,coloring,the number of neurons decreased significantly.Compared with the model group,the Balanophora polysaccharide treatment group is better than that of pathological changes in the model group,the Balanophora polysaccharide treatment group,cytoplasm disintegration is relatively small,relatively deep color,neurons can form,the number of neurons increased relatively.3 Immunohistochemical results: compared with sham operation group,model group in1,3,7 days after the three time in the spinal cord tissue LC3 protein positive cells increased significantly,with statistically significant difference(p<0.01).The Balanophora polysaccharide treatment group compared with model group,the high dose group after 1,3,7 day in the spinal cord tissue LC3 protein positive cells were higher than those in the model group(p<0.05),middle dose group in 3,7 days after operation was higher than in the model group(p<0.05);and,there was no significant difference between the high dose group(p>0.05),low dose group and model group had no difference(p>0.05).4 Western Blot results: 1 days after surgery,compared with sham group,the expression of Beclin in spinal cord tissue in the model group 1 protein significantly decreased,the expression of p62 and LC3-protein increased significantly,and the difference have statistical significance(p<0.01).The Balanophora polysaccharide treatment group compared with model group,the expression of Beclin 1and LC3-II protein in the spinal cord tissue of high dose group is higher than the model group(p<0.05),and the expression of p62 protein in the spinal cord tissue in the high dose group were lower than that of model group(p<0.05),and no significant dose and high dose differences between the two groups(p>0.05);there was no difference between the low dose group and model group(p>0.05).After 3 days,compared with the sham group,decreased expression of Beclin 1 protein in spinal cord tissue in the model group,the expression of p62 and LC3-II protein increased significantly,with statistical significance(p<0.01).Compared with the Balanophora polysaccharide treatment group and model group Relatively,low expression of Beclin 1 and LC3-II protein in spinal cord tissue of high dose group were higher than those in the model group,the difference was statistically significant in the low dose group(p<0.05),high dose group has statistical significance(p<0.01);the Balanophora polysaccharide,the expression of p62 protein in high dose group ridge spinal cord tissues were lower than those in the model group,the middle dose compared with the model group,the difference was statistically significant(p<0.05),high dose group has significant difference(p<0.01),low dose group and model group had no difference.The 7 day after surgery,compared with sham group,the expression of Beclin 1 and LC3-II protein in spinal cord tissue model in the group of decreased,the expression of p62 is increased,and the difference was significant statistical significance(p<0.01);the polysaccharide treatment group compared with model group,the low expression of Beclin 1 and LC3-II protein in spinal cord tissue of high dose group were higher than that of model group,In the dose group compared with model group,the difference was statistically significant(p<0.05),high dose group has significant difference(p<0.01),low dose group of LC3-II protein was also statistically significant(p<0.05);the Balanophora polysaccharide of low,high dose groups,expression of p62 protein in the spinal cord tissue were lower than the model group,low dose group compared with model group,the difference was statistically significant(p<0.05),high dose group has significant difference(p<0.01);and there were no significant dose and high dose differences between the two groups(p>0.05).Conclusion: Balanophora polysaccharide has protective effect on neurons after spinal cord injury,which may be related to the mechanism of enhancing Beclin 1 and LC3-II protein in spinal cord tissue and decreasing the expression of p62 protein and enhancing autophagy level. |
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