The Effect And Mechanism Of Kallistatin On Rheumatoid Arthritis

Rheumatoid arthritis(RA)is a kind of systemic chronic disease mainly composed of noninflammatory inflammation of joints and surrounding tissues.The main pathological features are chronic synovial membrane inflammation,thickening of lining layer,Inflammatory cell infiltration,formation of pannus,and destruction of cartilage and bone tissue,and then lead to joint deformity and loss of function,and finally can involve the serous,heart,lung and eye and other connective tissue extensive inflammatory disease.Tissue kallikrein(Kallistatin,Kal)is a serine protease inhibitor that binds specifically to Kallikrien(angiotensin)and inhibits its function.Kal has a variety of biological functions such as anti-inflammatory,anti-oxidation,inhibition of neovascularization,anti-tumor and so on.In this paper,according to the pathological features of RA and the biological function of Kal,mainly as the research object,from the in vitro cell experiments and in vivo animal experiments in both aspects of its anti-rheumatoid arthritis pharmacodynamic effects,and to explore its The mechanism of anti-rheumatoid arthritis.The main research contents of this subject include the following four aspects:1.Purification was carried out by using Pichia pastoris expression system,high density fermentation,centrifugation,salting out,hydrophobic chromatography and ultrafiltration.Then,the Kal protein was identified by Western blot,and the concentration of Kal was determined by ELISA.2.Chondrocytes were induced by type Ⅱ collagenase digestion,and induced by by IL-1 beta.3.The effect of Kal on chondrocyte inflammation model,respectively by(1)detecting the effect of Kal on cartilage cell toxicity and activity;(2)effects of Kal secretion of NO release inflammation of cartilage cells;(3)effects of Kal secretion of TNF-on inflammation of cartilage cells;(4)the influence of Kal on inflammation of chondrocyte apoptosis to investigate the effect of Kal on cartilage;cell inflammationmodel.The resμLts showed that the cartilage cells of Kal IL-1 beta induced inflammation model,can effectively protect the cell injury caused by IL-1 beta,mainly in: to improve chondrocyte viability;reduce the expression of inflammatory cytokines and inflammatory medium;through down regμLating the expression of apoptosis protein Bad,upregμLation of the expression of anti apoptosis protein Bcl-x L and apoptosis inhibition of cartilage cell.4.To investigate the effect of Kal on rat adjuvant rheumatoid arthritis model,it has been shown that in the animal model of rheumatoid arthritis,Kal can effectively inhibit the inflammatory response caused by complete Freund’s adjuvant,Adjuvant rheumatoid arthritis has a certain therapeutic effect,mainly in: Kal can reduce the CFA caused by knee swelling,reduce the expression of inflammatory mediators,inflammatory factors.In vitro cell experiments and in vivo animal experiments showed that Kal by reducing the secretion of inflammatory mediators and inflammatory factors,down-regulation of the expression of apoptotic protein Bad,up-regulation of anti-apoptotic protein Bcl-x L expression,inhibition of chondrocyte apoptosis,Arthritis play a therapeutic role.Indicating that Kal has a good anti-RA effect.