| Objective:Establishing an animal model of degeneration of the intervertebral disc,confirming the presence of nucleus pulposus-derived mesenchymal stem cells(NPMSC)in the degenerated intervertebral disc nucleus.And compare the biological characteristics of normal and degenerated intervertebral disc derived NPMSC.Comparison of Pleiotrophin(PTN)expression in normal and degenerated intervertebral disc-derived NPMSCs.To compare the effects of different concentrations of simvastatin on the expression of Pleiotrophin in MSCs with degenerated intervertebral disc and the biological characteristics of nucleus pulposus MSC.And explore the role of Pleiotrophin in this process.Methods:1.Making rat tail disc degeneration model:A disc degeneration model was established by percutaneous minimally invasive puncture of the caudal disc of the rat.2.Acquisition and culture of NPMSCs.3.Identification of NPMSC:Cell morphology observation,adherence growth,cell surface antigen protein molecular assay,three-line differentiation-inducing ability detection,dry gene expression.4.Comparison of biological characteristics of normal and degenerated intervertebral disc derived NPMSC.5.Effects of simvastatin on the biological function of degenerative intervertebral disc-derived NPMSCs and the expression of Pleiotrophin:Different concentrations of simvastatin were applied to degenerated intervertebral disc-derived NPMSCs to detect biological properties and PTN,BMP-2,aggrecan,type Ⅱ collagen gene expression.6.Effects of simvastatin on the biological properties of NPMSC overexpressing PTN gene:intervention of overexpression PTN gene NPMSC with simvastatin,and compared biological properties and the gene expression of PTN,BMP-2,aggrecan and type Ⅱ collagen.Results:1.We successfully using percutaneous minimally invasive puncture of the caudal disc of the rat to create a disc degeneration model.2.There are NPMSCs in the normal and degenerated intervertebral disc nucleus tissue.The biological properties of normal and degenerated intervertebral disc derived NPMSCs are different.The expression of PTN was found in degenerated intervertebral disc tissue and degenerated intervertebral disc derived NPMSC.The expression of PTN in degenerated intervertebral disc derived NPMSCs was significantly higher than that in normal intervertebral disc derived NPMSCs.3.Simvastatin can regulate the biological behavior of degenerated intervertebral disc derived NPMSC,and induce the expression of extracellular matrix:After simvastatin treatment,the expression of type II collagen and proteoglycan gene was found in all groups of NPMSCs,and its expression increased with the prolongation of action time.PTN and BMP-2 were present in each group of NPMSCs.The expression of PTN and BMP-2 in each group peaked on the 7th to 14th day after the action of simvastatin.The increase and decrease of PTN expression after simvastatin intervention is accompanied by an increase and decrease in the proliferative activity of NPMSC and a decrease and increase in apoptotic activity.4.NPMSCs biological activity and expression of PTN,BMP-2,proteoglycan and type Ⅱ collagen genes were enhanced after NPMSC transfected overexpressing PTN gene.Conclusion:Percutaneous minimally invasive puncture of the intervertebral disc anastomosis can successfully create a model of degenerated intervertebral disc.It was confirmed that the presence of NPMSC in the normal and degenerated intervertebral disc nucleus.It was confirmed that the difference in biological properties of normal and degenerated disc-derived NPMSCs.Confirming the expression of Pleiotrophin in degenerated intervertebral disc-derived NPMSCs and its effect on proliferation and apoptosis of NPMSCs.It was confirmed that simvastatin can regulate the expression of Pleiotrophin to regulate the biological properties of NPMSCs and promote extracellular matrix synthesis and expression.Simvastatin and Pleiotrophin can be used as a new direction for the treatment of intervertebral disc degeneration. |
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