Study On The Effects Of Panax Japonicus On Apoptosis-related Proteins After SCII Intervention Based On The Method Of Invigorating Qi And Activating Blood Circulation

ObjectiveBased on the method of Invigorating Qi and activating blood circulation,the preventive and therapeutic effects of Panax japonicus on spinal cord ischemia/reperfusion injury?SCII?in rats were studied.By comparing the histomorphological changes of motor neurons in the anterior horn of spinal cord after intervention with Panax japonicus,and detecting the apoptotic proteins,the protective effect of Tonifying Qi and activating blood circulation on motor neurons in the anterior horn of spinal cord after intervention with SCII was explored.Methods100 clean SD rats,male and female were randomly divided into sham-operated group,model group,low,medium and high dose group of Panax japonicus.After 7days of adaptive feeding,sham-operated group and model group were given 2 ml normal saline per day.The low,middle and high dose groups of Panax japonicus were given intragastric administration with concentration of 1.33g/kg.d^-1,2.67g/kg.d^-1,5.33g/kg.d^-11 respectively.The duration of intragastric administration was 7 days,and then the rats were made.The number and morphology of spinal motoneurons after SCII were observed by Nissl staining and HE staining,the content of neuron specific enolase?NSE?in serum was detected by ELISA,and the corresponding spinal motoneurons were detected by immunohistochemical staining.The expression of C-f o s,C-j u n and B ax proteins in the cells and the expression of C-f o s,C-j u n,B ax and B cl-2 proteins in the corresponding spinal cord tissues were detected by Western blot.Results1.Modified Ta r l o v Behavior Score:After SCII 6 and 24 hours,the scores of sham-operated group tended to be normal?p<0.05?;at the same time,compared with model group,the scores of middle-dose group were higher than model group?p<0.05?;compared with medium-dose group,the scores of low-dose group and high-dose group were lower than that of medium-dose group?p<0.05?.2.HE staining results:After SCII 6 and 24 hours,the morphology of spinal cord neurons in sham-operated group was intact,nucleolus and nuclear membrane were clear;the swelling of spinal cord neurons in model group was obvious,showing vacuole shape,and the cell body contracted seriously;the swelling of spinal cord neurons in low and middle dose groups of Panax japonicus decreased,and the nucleolus and nuclear membrane were clear,but the middle dose group was better than the low dose group.3.Nissl Staining results:After SCII 6 and 24 hours,the number of Nissl corpuscle granules in sham-operated group was relatively large,the morphology of neurons in model group was incomplete,and the cell gap was large,and the number of prominent decreased.The morphology of neurons and the number of Nissl body granules in the low and middle dose groups of Panax japonicus were significantly improved,but the middle dose group was better than the low dose group.4.ELISA results:After SCII 6 and 24 hours,the sham-operated group tended to be normal;at the same time,compared with the model group,the dosage group of Panax japonicus was lower than the model group?p<0.05?;compared with the middle dose group,the low and high dose group were higher than the middle dose group?p<0.05?.5.Immunohistochemistry results:The expression of C-f o s,C-j u n and B ax in spinal motoneurons of sham-operated group was basically the same after SCII 6 and24 hours?p<0.05?.At the same time,compared with model group,the expression of C-f o s,C-j u n and B ax in low and middle dose groups of Panax japonicus was down-regulated?p<0.05?,while the expression of C-f o s,C-j u n and B ax in high dose group was not significantly down-regulated?p<0.05?.Compared with the low dose group,the expression of C-f o s,C-j u n and B ax in the middle dose group decreased significantly?p<0.05?.6.Western Blotting results:After SCII 6 and 24 hours,the expression of C-f o s,C-j u n,B ax and B cl-2 in spinal cord neurons of sham-operated group was basically the same?p<0.05?;at the same time,compared with model group,the expression of C-f o s,C-j u n and B ax in middle-dose group of Panax japonicus was significantly down-regulated,and the expression of B cl-2 was up-regulated?P<0.05?.Conclusion:1.Invigorating Qi and Promoting Blood Circulation can relieve the damage of spinal cord neurons by interfering with SCII.2.Panax japonicus may reduce the excessive apoptosis of spinal cord neurons by inhibiting the expression of C-f o s,C-j u n and B ax proteins in spinal cord tissue after SCII and promoting the expression of B cl-2 protein.3.Based on the method of Invigorating Qi and activating blood circulation,the protective effect of Panax japonicus on spinal cord neurons after SCII may be achieved by inhibiting excessive apoptosis.