Experimental Study On The Regulation Of Neurocyte Apoptosis By Longzhong Xiaozhong Zhitong Mixture Based On JAK2/STAT3 Signal Pathway In Rats With Spinal Cord Injury

Objective:Based on previous studies in which the JAK2 /STAT3 signal pathway was used as the target of Longzhong Xiaozhong Zhitong Mixture in the treatment of SCI,a spinal cord injury model was successfully constructed.After the intervention of Longzhong Xiaozhong Zhitong Mixture,the expression of its related proteins in spinal cord tissue was detected,and the mechanism of Longzhong Xiaozhong Zhitong Mixture in regulating neuronal apoptosis in SCI rats was explored,Provide a certain theoretical basis for molecular biology research.Methods:Select 96 adult male SD rats and randomly divide them into normal group,The sham surgery group,model group,Longzhong Xiaozhong Zhitong Mixture group,sodium aescinat group,and Longzhong Xiaozhong Zhitong Mixture+AG490 inhibitor group.The SCI animal experiment model was established using the modified Allen method.After the model was successfully established,the drug was administered by gavage after surgery.The normal group,The sham surgery group and the model group were given physiological saline of 2 m L /(day *time),respectively.The Longzhong Xiaozhong Zhitong Mixture group was given Longzhong Xiaozhong Zhitong Mixture of 1.7 m L /time,once each day in the morning and evening;Longzhong Xiaozhong Zhitong Mixture+AG490 Inhibitor Group: Longzhong Xiaozhong Zhitong Mixture was administered orally 1.7m L/time,once in the morning and once in the evening each day,while daily intraperitoneal injection of AG490 inhibitor 15 mg /Kg was performed.In the sodium aescinat group,1.35mg/kg sodium aescinat was administered orally twice a day.According to the BBB(Basso,Beattie&Bresnahan locomotor rating scale,BBB)scoring standard,the motor function of the hind limbs of rats was observed and recorded on the 1st,3rd,7th,and 14 th days after surgery.HE staining method was used to observe the histological morphology of the spinal cord of rats in each group at 1,3,7,and 14 days after surgery;Western blot method was used to detect the changes of protein expression levels of p-JAK2,p-STAT3,and Caspase-3 in spinal cord tissue of normal group,model group,Longzhong Xiaozhong Zhitong Mixture group,sodium aescinate group,and Longzhong Xiaozhong Zhitong Mixture+AG490 inhibitor group at 3 days after surgery;Immunohistochemical method was used to detect the expression level of caspase-3 positive cells in the spinal cord tissue of the normal group,the model group,the Longzhong Xiaozhong Zhitong Mixture group,the sodium aescinate group,and the Longzhong Xiaozhong Zhitong Mixture+AG490 inhibitor group at 3 days after surgery.Relevant data were obtained and analyzed.Results:1.After SCI,the functional activity of both lower limbs of SD rats was abnormal,and the BBB score was significantly decreased(P<0.05).HE staining showed that the morphology of spinal cord tissue changed after SCI.Intervention with Longzhong Xiaozhong Zhitong Mixture and sodium aescinat could improve the BBB score,with no statistical difference between the two groups(P>0.05).At14 days,the BBB score of Longzhong Xiaozhong Zhitong Mixture was significantly higher than that of sodium aescinat group,with statistical significance(P<0.05).2.After spinal cord injury,the expression of P-JAK2,P-STAT3,and Caspase-3in spinal cord tissue increased,which was positively correlated with the degree of SCI.After the intervention of Longzhong Xiaozhong Zhitong Mixture,the expression of P-JAK2,P-STAT3,and Caspase-3 in spinal cord tissue decreased,neuronal apoptosis decreased,and the neural function of rats gradually recovered.3.Compared with Longzhong Xiaozhong Zhitong Mixture,the expression of PJAK2,P-STAT3,and Caspase-3 in sodium aescinat was not statistically significant(P>0.05);Compared with the Longzhong Xiaozhong Zhitong Mixture group,the expression levels of P-JAK2,P-STAT3,and Caspase-3 in the spinal cord tissue of the Longzhong Xiaozhong Zhitong Mixture+AG490 inhibitor group were lower,with statistical significance(P<0.05).Conclusion:1.Both Longzhong Xiaozhong Zhitong Mixture and sodium aescinat can promote the recovery of lower limb function in SD rats,and their long-term efficacy is better than that of the sodium aescinat group.2.After SCI in rats,the JAK2/STAT3 signaling pathway is abnormally activated,promoting neuronal apoptosis,which may have a regulatory effect on secondary spinal cord injury.3.Longzhong Xiaozhong Zhitong Mixture and sodium aescinate can reduce the expression level of Caspase-3 protein and reduce neuronal apoptosis caused by secondary spinal cord injury.The mechanism of action may be related to the inhibition of the activation of the JAK2/STAT3 signaling pathway.