Mechanism Of Buyang Huanwutang Combined With BMSCs Transplantation In Treatment Of Spinal Cord Injury Based On PI3K/AKT Signaling Pathway

Objective: To investigate the effect of Buyang Huanwu Decoction combined with bone marrow mesenchymal stem cells（BMSCs）transplantation on rats with spinal cord injury（SCI）and its mechanism,as well as the effect of Buyang Huanwu Decoction on the migration of BMSCs in rats,and discuss its mechanism.Methods: 60 female SD rats were used,50 of them were randomly selected to prepare SCI rat model by modified Allen heavy object method,and the remaining 10 were used as sham operation group.All SD rats were divided into 6 groups,including sham operation group,model group,Buyang Huanwu Decoction group,BMSCs transplantation group,Buyang Huanwu Decoction+BMSCs group,Buyang Huanwu Decoction+BMSCs+LY294002 group,with 10 rats in each group.The sham operation group was given the same amount of normal saline every day,the model group was given the same amount of normal saline every day,the Buyang Huanwu Decoction group was given the same amount of Buyang Huanwu Decoction(25g·kg^-1),the BMSCs transplantation group was given 1m L BMSCs through the tail vein,and the Buyang Huanwu Decoction+BMSCs group was given the same amount of Buyang Huanwu Decoction(25g·kg^-1)through the tail vein Buyang Huanwu Decoction+BMSCs+LY294002 group was given Buyang Huanwu Decoction(25g·kg^-1)by gavage and 1m L BMSCs and LY294002(40mg·kg^-1)by tail vein.The recovery of neuromotor function in rats was observed through modified Tarlov scores and oblique plate tests at 1w,3w,and 5w.Immunohistochemical staining was used to detect the number of positive cells labeled with 5-bromo-2-deoxyuracil（Brdu）in spinal cord tissue,Western blot was used to detect the expression of phosphorylated protein kinase B（p-Akt）,glycoprotein 130（gp130）and interleukin-6（IL-6）in spinal cord tissue.Result:1.Tarlov score: Compared with the model group,Tarlov score of sham operation group and each treatment group increased significantly at 1w,3w and 5w（P<0.05）.Compared with BMSCs group,the Tarlov score of Buyang Huanwu Decoction+BMSCs group increased significantly at 1w,3w and 5w（P<0.05）.Compared with Buyang Huanwu Decoction+BMSCs group,the score of Buyang Huanwu Decoction+BMSCs+LY294002 group increased significantly at 1w,3w and5w（P<0.05）2.Inclined plate test: Compared with the model group,the critical angle of the inclined plane in the sham operation group and each treatment group was significantly higher at 1w,3w and 5w（P<0.05）.Compared with the BMSCs group,the critical angle of tilt plane in the Buyang Huanwu Decoction+BMSCs group was significantly higher after the combination of Buyang Huanwu Decoction and BMSCs transplantation（P<0.05）.Compared with Buyang Huanwu Decoction+BMSCs group,the critical angle of tilt plane in Buyang Huanwu Decoction+BMSCs+LY294002group was significantly higher（P<0.05）.3.Immunohistochemical staining results: There were no Brdu labeled positive cells in spinal cord tissue of sham operation group.Brown and yellow Brdu labeled positive cells were found in spinal cord tissue of BMSCs transplantation groups.At 5w of transplantation,compared with BMSCs group,the number of Brdu positive cells in Buyang Huanwu Decoction+BMSCs group increased significantly（P<0.05）.Compared with Buyang Huanwu Decoction+BMSCs group,the number of Brdu positive cells in Buyang Huanwu Decoction+BMSCs+LY294002 group increased significantly（P<0.05）.4.Western blot test results: There was a significant difference in the expression of spinal cord tissue related proteins between the sham operation group and the model groups of each treatment group（P<0.05）.Compared with the sham operation group,the expression levels of p-Akt,gp130 and IL-6 protein in the spinal cord tissue of the model group were significantly increased（P<0.05）.Compared with the model group,the expression levels of p-Akt,gp130 and IL-6 protein in the spinal cord of rats in the sham operation group and each treatment group were significantly decreased（P<0.05）.Compared with BMSCs group,the expression levels of p-Akt,gp130 and IL-6 protein in Buyang Huanwu Decoction+BMSCs group were significantly decreased（P<0.05）.Compared with Buyang Huanwu Decoction+BMSCs group,the expression levels of p-Akt,gp130 and IL-6 protein in Buyang Huanwu Decoction+BMSCs+LY294002 group were significantly decreased（P<0.05）.Conclusions: 1.Buyang Huanwu Decoction and BMSCs transplantation can effectively improve the motor function of SCI rats,and the combination of the two has a synergistic effect.2.Buyang Huanwu Decoction promotes the migration of BMSCs and the recovery of spinal cord function by inhibiting PI3K/Akt signal pathway.3.Buyang Huanwu Decoction combined with BMSCs can alleviate the inflammatory reaction in the injured area by inhibiting the PI3K/Akt signal pathway,thus promoting the recovery of spinal cord function.