Study On The Expression Of MMP-2,MMP-9 And TIMP-1 Rheumatoid Arthritis

Rheumatoid arthritis is a kind of chronic general auto-immuned disease which ischaractered with synovitis in joints, with hyperplasia and destructive lesions which islike "local malignant tumors", developed progressively. In the late stage, thedestruction of joint cartilage and bone leads to joint abnormity and dysfunction. It'sincidence is about 0.34-0.36% in our country. However, the mechanism has not beenexplained.Recently, many researchers think that there is a close link between matrixmetalloproteinase and rheumatoid arthritis. MMPs is a family of protein-enzyme withZn~+ as cofactor. It's main fuction is to degrade the extracellular matrix. And it is one ofmost important enzyme series on regulate the dynamic equilibrium of ECM. MMPscan degrade the collagen, proteoglycan and other big molecule of ECM in the jointcartilage and bone, and it also can facilitate the invasion of the pannus tocartilage. That is the main resources of injury in RA from the invasion of theproliferative pannus to cartilage of joint. Finally, the systematic proliferative andinvasive synovitis produced. Up to now, there have been over 20 kinds of MMPs, andMMP-2 and MMP-9 belong to gelatinase. They can resolve gelatin, typeⅠ, typeⅣ,typeⅤ, typeⅨcollagen and conglutinin. They are the important components in thejoint cartilage, and they have a relationship with RA. Tissue matrix metalloproteinaseinhibitor(TIMPs) is a natural inhibitor of MMPs and is a group of factors withmultifunction which can inhibit the activity of MMPs. They can inhibit activitives ofMMPs in the many links to prevent the degradation of ECM. And in RA, in order tosustain the balance of the ECM, TIMPs can make some changes in expressional levels.There are 4 kinds of TIMPs, TIMP-1 can inhibit most of the MMPs, but it is mainly the specific inhibitor of MMP-9, and it can form a complex with high affinity andnoncovalent bond. The Expression of MMP-2,MMP-9 and TIMP-1 in rheumatoidarthritis has being studied.Objective:To detect the expression of the MMP-2, MMP-9 and TIMP-1 in the lining cell ofsynovium and subsynovial tissue of rheumatoid arthritis and to evaluate the role theyplay in the mechanism of RA.Material and Methods:1. In this retrospective study, immunohistochemical staining for MMP-2, MMP-9and TIMP-1 is performed in 65 cases of RA in Tianjin hospital(from 2004 Jan to 2006Oct). Among these cases, 43 are female, age range from 17 to 70, and average is 37±0.12 years old, 34 occur in knee joints, 2 in hip joints, 3 in elbows joints, 3 in anklejoints, and 1 in wrist joint. There are 22 male, age range 12 to 71, average is 39±0.10 years old, 17 occur in knee joints, 2 in hip joints, 2 in ankle joints, and 1 inwrist joint. Besides, 10 cases OA and 7 cases synovium from meniscus injury arechosen as control group. Among 7 cases, patients who are chosen are young group,age range is 11 to 35 years old, average is 22±0.20, they are operated within 3 daysfrom injured.2. 65 cases mentioned above which are fixed by Formalin, embedded by paraffinare performed using immunohistochemical stain to detect the expression of MMP-2,MMP-9 and TIMP-1 respectively.3. Biologic light microscopes is used to recognize and calculate the quantity ofpositive cells of MMP-2, MMP-9 and TIMP-1. Select 10 fileds of vision randomly andcount positive cells per filed of vision, calculate the sum number/10, and regard "=10"as negative; regard "11～25" as "+"; regard "26～50" as (++); regard "=51" as"+++". When recongnize and calculate the number of positive cells of MMP-2,MMP-9 and TIMP-1. And as standard like above, count the number of positive cells inthe lining cells and subsynovial tissue.4. Data processing: analyse the datum by SPSS 11.5. Rank sum test is used to analyse the expression of MMP-2, MMP-9 and TIMP-1 in RA, OA and approximatelynormal synovium. Rank sum test is used to analyse the expression of MMP-2, MMP-9and TIMP-1 between lining cells and subsynovial tissue. Spearman rank correlationtest is used to evaluate the correlation of expression among MMP-2, MMP-9 andTIMP-1.Results:1. The expressional rates of MMP-2 in RA, OA and approximate normal synoviumare 66.15%(43/65), 20.00%(2/10), 14.29%(1/7) respectively; The expressional ratesof MMP-9 in RA, OA and approximately normal synovium are 60.00%(39/65),10%(1/10), 0.00%(0/7) respectively; The expressional rates of TIMP-1 in RA, OAand approximate normal synovium are 73.85%(48/65), 30.00%(3/10), 0.00%(0/7)respectively. In a word, the positive expressional rates of MMM-2, MMP-9 andTIMP-1 are higher than the ones of OA and approximate normal synovium noticeably(p＜0.05).2. The expressional rate of MMP-2 in lining cell of RA is 40.00%(26/65), and theone of it in the subsynovial tissue of RA is 61.54%(40/65), and the latter is higher thanthe former noticeblely(p＜0.05). The expressional rate of MMP-9 in lining cell of RAis 69.23%(45/65), and the one of it in the subsynovial tissue of RA is53.85%(35/65),and the former is higher than the latter noticeblely(p＜0.05). The expressional rate ofTIMP-1 in lining cell of RA is 60.00%(39/65), and the one of it in the subsynovialtissue of RA is 35.38%(23/65), and the former is higher than the latter noticeblely(p＜0.05).3. By Spearman rank correlation test, the value of p between MMP-2 and MMP-9is more than 0.05, that is the two have noncorrelation. By Spearman rank correlationtest, the value ofp between MMP-9 and TIMP-1 is less than 0.05, that is the two havenoticeable correlation. By Spearman rank correlation test, the value of p betweenMMP-2 and TIMP-1 is less than 0.05, that is the two have noncorrelation.Conclusion:1. The positive expressive rates of MMP-2, MMP-9 and TIMP-1 are higher than OA and approximate normal synovium noticeablely. And deduce that they have a rolein the mechanism of RA.2. MMP-2, MMP-9 and TIMP-1 have different positive expressional rates insynovium.and subsynovial tissue of RA. And deduce that MMPs and TIMPs havedifferent expressive rates.3. There are strong correlation between the positive rates of MMP-1 and TIMP-1,because TIMP-1 is the specific inhibitor of MMP-9, and TIMP-1 can prevent thedegradation of ECM from MMP-9.