TGF-Beta 1, MMP-14,P38 MAPK Influence On Secondary Frozen Shoulder And Mechanism Research

Objective the study through arthroscopy acquire synovial membrane under acromion in patients with secondary frozen shoulder;the original generation cultivation of synovial fibroblasts under acromion,make frozen shoulder animal model of rats,to explore the pathogenesis of secondary frozen shoulder pathological mechanism,and further explore the influence and function mechanism of TGF-beta 1,MMP-14,and MAPK pathway to the secondary frozen shoulder patients’ s shoulder peak synovial hyperplasia,vascular formation.What’s more,for the secondary prevention and control of frozen shoulder open up new ideas and new ways.Methods 1.By arthroscopy to acquire secondary frozen shoulder patients and normal patients’ synovial under acromion before operation.By Western Blot,and immunohistochemistry,PCR technique to detect the protein and gene(TGF-beta 1,MMP – 14,P38 MAPK,VEGF)expression in synovial under acromion of patients with secondary frozen shoulder and normal.Preliminary discussion of the influence and function mechanism on related genes to the synovial proliferation and angiogenesis under acromion;2.cultured the original generation fibroblasts under acromion,through the TGF-beta 1 cytokines induced primitive cells by different concentration gradient;Western Blot,and immune fluorescence PCR technique was used to detect MMP-14,P38 MAPK,VEGF protein and gene expression;further explored the influence and function mechanism of related genes to the synovial proliferation and angiogenesis under acromion;3.Artificially to injury rats rotator cuff making secondary frozen shoulder model,and after 12 weeks by Western Blot,PCR technique to detect related proteins and gene expression around the shoulder joint synovial tissue;further verify the influence and function mechanism of related genes to the synovial proliferation and angiogenesis under acromion.Results 1.We could find the synovial proliferation,angiogenesis under acromion significantly in patients with frozen shoulder to the normal.The TGF-beta 1,MMP-14,P-P38 MAPK,VEGF related gene expression was increased in the synovial tissue under acromion in patients with frozen shoulder.2.Primitive fibroblast reach steady state after 2 to 3 generations,through the TGF-beta 1 cytokines different concentration gradient(0,5,10,20 ng/ml)induced primary cell,as the concentration of the faster increase fibroblast proliferation.Western Blot,PCR detection shows that p-p38,MMP-14,vegf gene expression increased with the increase of TGF-beta 1 concentration.3.The TGF –beta 1,p-p38,MMP – 14,vegf gene expression increased significantly in synovial tissue of Secondary frozen shoulde rats injury side to the normal side,with the passage of time the more obvious increase trend.Conclusion 1.Secondary frozen shoulder patients’ synovial hyperplasia,vascular formation increase under acromion is the significant cause of shoulder joint tissue fibrosis and limited shoulder joint activity.2.TGF-beta 1,MMP-14 and MAPK pathways have a certain relationship to the synovial hyperplasia,increased blood vessel formation under acromion in patients with frozen shoulder.TGF-beta 1 through the induction of MAPK pathway in P-P38,so that the shoulder synovial fibroblasts proliferation,adhesion,organized into fibrosis hypoxia inflammation,vascular endothelial cell proliferation,angiogenesis,causing symptoms of frozen shoulder;on the other hand TGF-beta 1can raise MMP-14,cause disorder of MMP/Timp balance,promote fibroblasts proliferation,angiogenesis,tissue fibrosis adhesion,further resulting in secondary frozen shoulder symptoms.