The Effects Of Jishen Bitong Formula Combined With Bone Marrow Mesenchymal Stem Cells Transplantation On Degenerative Intervertebral Disc Repair And Expression Of CXCR4 And NF-?B

Objective:In vitro culture of BMSCs by whole bone marrow wall-sticking method.To establish the model of rat tail intervertebral disc degeneration and to observe the effects of Jishen Bitong formula combined with BMSCs transplantation on degenerative intervertebral disc repair and expression of CXCR4 and NF-?B.Methods:BMSCs was extracted from rat femur bone marrow,the whole bone marrow wall-sticking method was cultured in vitro to P3 generation,and was identified by morphological observation,cell surface marker detection,proliferation observation,and cryopreservation in liquid nitrogen for subsequent transplantation.The model of rat tail intervertebral disc degeneration was fabricated by percutaneous fibrous annular puncture,and its degeneration was detected dynamically by magnetic resonance imaging.50 model rats with the same degree of degeneration were selected randomly into 5 groups.The specific intervention methods are:Group A:BMSCs Transplantation of rat tail vein and high dose "Jishen Bitong formula" decoction for gastric lavage;Group B:BMSCs Transplantation of rat tail vein and medium dose "Jishen Bitong formula" decoction for gastric lavage;Group C:BMSCs Transplantation of rat tail vein and low dose "Jishen Bitong formula" decoction for gastric lavage;Group D:BMSCs Transplantation of rat tail vein and the equal volume pure water for gastric lavage;Group E:PBS liquid injection of rat tail vein and the equal volume pure water for gastric lavage.After 1 months of intervention,MRI was reviewed and HE staining was combined to observe the restoration of degenerative intervertebral discs.The expression of CXCR4 and NF-?B in serum and intervertebral discs was detected by ELISA.Results:In vitro culture of rat P3 generation BMSCs by whole bone marrow wall-sticking method,and the morphological observation showed that the structure uniformity was higher and the proliferative ability was stronger.Through flow cytometry,the classical cell surface markers were identified as BMSCs.Most of the degenerative disc degeneration in rats made by percutaneous fibrous annular puncture is improved Thompson Grade ?.After 1 months of treatment,from MRI and HE staining,it was found that Jishen Bitong formula combined with BMSCs transplantation can effectively delay the degeneration of rat tail intervertebral disc,the results were better than those of simple BMSCs transplantation.There were CXCR4 and NF-?B expression in the intervertebral discs and serum of each group.The contents of CXCR4 and NF-?B in the same group were significantly higher than those in the intervertebral discs(p<0.05),and the difference was statistically significant.The CXCR4 and NF-?B of the A-D group were significantly higher than those in the blank control group(p<0.05),and the difference was statistically significant.The high dose group(Group A),the medium dose group(Group B)and the low dose group(Group C)can promote the expression of CXCR4 and NF-?B,and have a concentration dependence.Conclusion:The whole bone marrow wall-sticking method can produce P3 generation BMSCs with high purity and good proliferation.The model of rat tail intervertebral disc degeneration made by percutaneous fibrous annular puncture was simple,small damage,high success rate and feasibility.MRI has the advantages of safety,no radiation,no bone artifact and high contrast,and can be used to detect intervertebral disc degeneration related diseases in animals.The degree of intervertebral disc degeneration can be evaluated objectively with the help of tissue staining technology.BMSCs transplantation can delay intervertebral disc degeneration,combined Jishen Bitong formula can improve its repair ability,and the mechanism of Jishen Bitong formula combined with BMSCs transplantation to repair intervertebral disc degeneration may be related to the increase of CXCR4 and NF-?B expression.