Concentration Of IL18 And Other Cytokines In Synovial Fluid And Chondrocytes In Patients With Osteoarthritis And The Significance

BackgroundsOsteoarthritis(OA) is one of the four biggest senile diseases of our country,and make the older disable.The pathogenesis isn't clear up to now.The current research show that the pathogenesis of OA is subjected to regulation of various genes,involves kinds of cytokines,which form cascade network system,make chondrocytes cluster, cartilage matrix degrade,and result in vicious cycle.The expression of cytokines of matrix metalloproteinase(MMPs) and interleukins(ILs) is more conspicuous in OA chondrocytes comparing with normal chondrocytes by immunohistochemistry. Interleukin-18(Interleukin-18,IL-18) are cloned from the toxic shock mouse' liver by Okamura in 1995,it is the protein induced by the stress,The protein is peptide fragment with the molecular weight of 18-19ku,because the peptide can induce helper T lymphocytes produce interferon-γ(IFN-γ),it can enhance the ability of splenic T cell proliferation and splenic NK cell activity,and interleukin-12(IL-12) the biological activity very similar,it was originally named IFN-γinducible factor (IFN-γIGIF).Current study showed that,IL-18 is not only found in synovial fluid, synovium,meniscus,cartilage cells of patients with arthritis,also be found in the serum,but the content is lower than in synovial fluid.Gracie found the IL-18 content of synovial fluid and synovial membrane of rheumatoid arthritis higher than osteoarthritis,and the osteoarthritis patients' content was higher than normal,but the expression level of IL-18 was closely related to and activity period of osteoarthritis,this prompted us IL-18 played an important role in the occurrence and development of osteoarthritis,it may be involved in the degradation of cartilage matrix,cartilage and synovial cell apoptosis of the inflammatory lesion process. IL-18 in synovial fluid direct contact T lymphocytes,increased monocytes produced TNF-αand IL-1β,IFN-γinduced by IL-18 in the synovial fluid can significantly stimulate IL-6,NO,IL-β,PGE2,and IFN-γ,and TNF-αinduced inflammatory diseases,including the occurrence and development of arthritis.Studies of levels of IL-18 expression in synovial tissue with arthritis have shown its expression have a positive correlation with IL-1,TNF-α,local synovitis level,and in parallel to the acute phase of disease,suggesting that IL-18 is a important early of cytokine arthritis. The cytokines invovled in OA pathology don's work by single way but interacting,form cascade reaction.So it is important to study kinds of cytokines expression simultaneously.The application of proteinome array technique provide convenient instrument to study the much cytokines expression simultaneously. Liquichip workstation system is a new platform to proteinome study.Transversal compareing these cytokines relationship,help to find out the key cytokine that result in OA.Liquid-chip,xMAP(flexible Multi-Analyte Profiling) technology to carry out substantial factor test to study the expression of IL-18 binding protein to other pathogenic factor expression in order to study IL-18 and other factors the synergy between as well as the mutual influence.It can help to approaching mechanism of IL-18 in pathology,it can provide more scientific basis of clinical applications.Objectives1.Study the expression of IL-18 and prostaglandin E2 and other cytokine in the pathogenesis of OA in synovial fluid of the patients with OA,and interleukin-18 influence other growth factor and cytokine' expression,find out IL-18 in the OA disease process and to analyze the differences of their expression in normal articular organize.And to observe the changes of IL-18 and prostaglandin E2 expression in the time and space with interleukin-18 binding protein influence,in order to explore IL-18 binding protein in the process of the molecular mechanism of OA,fine out the new ways of OA treatment diagnosis and treatment.2.To get wareness of the morphological changes a of the cartilage cells in the passage. Make a understand of the difference between normal and abnormal in morphology.3.Study the changes of expression of IL-18 and prostaglandin E2 in the pathogenesis of OA in articular cartilage cells of the patients with OA,and interleukin-18 influence other growth factor and cytokine' expression,find out IL-18 in the OA disease process and to analyze the differences of their expression in normal articular organize.And to observe the changes of IL-18 and prostaglandin E2 expression in the time and space with interleukin-18 binding protein influence,in order to explore IL-18 binding protein in the process of the molecular mechanism of OA,fine out the new ways of OA treatment diagnosis and treatment.Methods1.Culturing normal chondrocytes and OA chondrocytes(from the patients with TKA). Separation of cartilage cells by the Enzymatic digestion,subcultured and observation the differences in Morphological2.Detecting the cytokines(PGE2,IL-6,8,TGF-βand VEGF) in the supernatants by liquichip workstation,and find out the key cytokines.Comparing the diffenert cytokines between them,find out the key cytokines,and approach the effect of IL-18 on the cytokines to understand its application in OA pathology.3.Data presented by(?)±S,statistics done with SPSS13.0.Inter-group comparison group using t-test,use correlation and linear regression analysis between IL-18 and related factors.P＜0.05 presents statistics different.Results1.Synovial fluid of OA patients with PGE2,IL-6 and IL-8 levels have increased follow the IL-18.Correlation analysis showed that between IL-18 and PGE2,IL-6 and IL-8 has a positive correlation.Studies shows that PGE2,IL-6 and IL-8 these pathogenic factor play a very important role in OA.They are important pathogenic factor,And IL-18 can significantly enhance the expression of factor of these cytokines in synovial fluid of the OA patients.2.There isn's different in morphology between the normal chondrocytes and OA chondrocytes.The time of OA chondrocytes in the form of adherence and primary culture is longer than the normal chondrocytes.The chondrocytes occur dedifferentiation after passage 4.The normal chondrocytes' body is round,nucleus is round or ellipse,coloration of nucleus is moderate,there are bulks of dilated rough endoplasmic reticulum(RER) around nucleus,more glycogenosome lie in intracytoplasm.The OA chondrocytes' body is small,with much reductus.The nucleus is larger and its coloration is deeper,with polygon because of indentation.Less RER and glycogenosome lie in intracytoplasm.Cytoplasm's coloretue of the postive chondrocytes(with one-antibody) is deeper,with brunneus. The negative(without one-antibody) is tasteless.3.The level of PGE2,IL-6,8 and IL-18 in OA chondrocytes is higher than in normal chondrocytes.Chondrocytes of OA patients with PGE2,IL-6 and IL-8 levels have increased follow the IL-18.Correlation analysis showed that between IL-18 and PGE2,IL-6 and IL-8 has a positive correlation.Studies shows that PGE2,IL-6 and IL-8 these pathogenic factor play a very important role in OA.They are important pathogenic factor,And IL-18 can significantly enhance the expression of factor of these cytokines in chondrocytes of the OA patients.4.Liquichip workstation liquid protein chip system detects six kinds of the expression of cytokines and found that OA chondrocytes express IL-18 expression high,PGE2, IL-6 and IL-8 level was significantly higher than that of normal cartilage cells level,Conclusions1.The morphology different between normal chondrocytes and OA chondrocytes aren't conspicuous,but the funtion of OA chondrocytes in synthesing protein and glycogenosome decrease by transmission electron(TEM).2.The level of PGE2,IL-6,8 and IL-18 in OA chondrocytes is higher in OA than in normal chondrocytes.3.Cytokines PGE2,IL-6 and IL-8 etc.play the role of pro-inflammatory in the pathological process of OA to,it may be involved in articular cartilage damage,and promote the progress of OA,IL-18 can significantly enhance expression of PGE2, IL-6 and IL-8 in synovial fluid and articular cartilage cells,indicating that IL-18 may be involved in articular cartilage damage,and promote the progress of OA,If you can inhibit the expression of IL-18 is an effective way to slow down the pathological process of OA.IL-18 inhibitors can inhibit the expression of IL-18 can exert its protective role in cartilage.4.The Liquichip workstation is a new detecting system with high-flux and hypersensitivity,help to study proteome.