| Objectives: 1.Observe the expression of autophagy after spinal cord injury in rats and the expression changes at different time points,new ideas were provided for the study of secondary injury of spinal cord injury.2.Observe the effect of hydrogen-rich water on the functional recovery of rats after spinal cord injury and the effect of autophagy expression in spinal cord tissue,the possible mechanism of hydrogen-rich water in the treatment of spinal cord injury was preliminarily discussed.Methods: fifty female SD rats with body weight of 260 30 g were randomly divided into three groups: the sham group(n = 10),the control group(n = 20),and the experimental group(n = 20).In the sham operation group,only t11-t12 lamina was removed,but the spinal cord was not injured.The rats in the control group were treated with modified Allen blow method to establish a spinal cord injury model.Rats in the experimental group were injected with hydrogen-rich water containing 160 ppm per day once a day after the establishment of spinal cord injury model by the modified Allen blow method.After the successful establishment of the model,the drug was administered for 14 consecutive days after the operation,and the BBB score of each group of rats was recorded daily.The expressions of autophagy related proteins LC3 I,LC3II and Beclin1 in rats were detected by Western blot at 1d,3d,7d and 14 d after surgery,and the distribution and expression of Beclin1 in autophagy proteins in rats were detected by immunohistochemistry,and the ultrastructure of nerve cells and the formation of autophagosomes were observed under transmission electron microscope.Results: 1.The experimental group's 3d BBB score was slightly higher than that of the control group at 1d and 3d after surgery,and its 14 d BBB score was significantly higher than that of the control group at 7d and 14 d BBB after surgery,with statistically significant difference(p < 0.05).2.Western blot results of rat spinal cord tissues showed that Beclin1 expression and LC3II/LC3 values increased after spinal cord injury compared with the sham operation group.Beclin1 expression and LC3II/LC3 values in the experimental group were lower than those in the control group at each time point,and the difference was statistically significant(p < 0.05).3.Immunohistochemical results showed that Beclin1 protein was expressed in the cytoplasm,and the average optical density value of the protein was calculated by Image Pro Plus 6.0 software.It was found that the average optical density value of the protein in the experimental group at each time point was lower than that in the control group,and the difference was statistically significant(p < 0.05).4.In the result of tem were microscopically control basic normal nerve membrane and organelles,autophagy rare,and the experimental group and control group 1 d,3 d spinal cord tissue after injury is to a large number of autophagy,organelles swelling damage is serious,the 7 d,14 d after two groups of the structure of the survival of the nerve cells gradually returned to normal,but the rich hydrogen water complex is better than that of saline group.Conclusion: 1.Under normal conditions,autophagy is expressed at low levels in the spinal cord tissue of rats to maintain cell homeostasis.After spinal cord injury,the expression of autophagy related proteins in the spinal cord was up-regulated and reached a peak 3 days later,and then decreased.The expression was still high 14 days after the surgery,indicating that autophagy overexpression after spinal cord injury caused autophagy death,which may be one of the important mechanisms of secondary spinal cord injury.2.Hydrogen-rich water treatment can improve the recovery of hind limb function after spinal cord injury in rats,and down-regulate the expression of autophagy related proteins to reduce the autophagy death of cells,which plays a protective role in spinal cord injury,indicating that the down-regulation of autophagy in spinal cord injury is the mechanism of hydrogen-rich water treatment for spinal cord injury. |
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