Study Of The Expression Of Matrix Metalloproteinases(MMP-2,9) And Their Tissue Inhibitors(TIMP-1,2) In Endometriosis And Its Correlation To The Invasion And Metastasis

Endometriosis(EM) is a common gynaecological disease.There is anincreased morbidity recently.Its recurernce rate in five years is morethan 40%, regardless of whatever treatment, and it mainly due to itspathogenesis has not yet clear. Although EM is a benign disease, but itscharacteristics of hyperplasia, excitement, extensive cultivation andeasily relapse are similar to malignant neoplasms,named A Benign Cancer,so it is dificult to cure.Matrix metalloproteinases (MMP) familyare important enzymes in degradation process of extracellularmatrix . Tissue matrix metalloproteinases (TIMP) are inhibitors of MMP.Research confirmed : the balance of MMP and Timp extracellular matrixis relative to various pathological conditions, especially with theinvision and transformation of tumour. Many studies have shown MMPs(MMP-2, 9) and Timp(TIMP-1, 2) play an important role in the pathogenesisof EM. The pathogenesis of EM contains the development of adhes0n ,invision and vascularization of ectopic endomembrane.So accommodationor intervesion of these processes may be new laboratorial and academicbasis on treatment. Anti-invision treatment and supressing act ivity ofMMPs are new target for curing EM.My paper studied MMP-2,9andTIMP-1,2 in sera, ascites and tissuesof patients with endometriosis levels from gene to protein,andtheir characterization, positioning, quantitative detection byimmunohistochemical method, gelatin zymogram analysis and RT-PCR .Thenstudied the interaction between MMP-2, 9 and TIMP-1,2 and the importantfunction in the invision and transformation process of EM .So MMP-2,MMP-9 are further confirmed as a better molecule marker for EM, and isexpected to be a new special marker in diagnosis.Now the main treatment contains endocrine and surgery ,which arenot satisfactory. Anti-invision therapy will become a new method. Myresearch studied the method of using external pyrogallic acid to treatrats with EM ,which were body transplanted. Then studied the variationsof size, pathological changes and ultrastructure by light and electronmicroscope and levels of MMP-2,9 and TIMP-1,2 of sera andtissues by immunohistochemical method, gelatin zymograrn analysisbefore and after therapy.We also test hormone levels by radio-immunoassayand make known of the effect of pyrogallic acid on the cultivation andgrowth of ectopic endowmembrane ,then discuss the foregroud of externalMMPs in treating EM.The results of immunohistochemical method are: there is highexpression of MMP-2,9 in the ectopic endomembrane than normal andreigning endomembrane and TIMP-1,2 are significantly lower than normaland reigning endomembrane (P<0.01).However, there is no statistcaldifference of MMP-2,9 and TiMP-t. 2 between normal and reigningendomembrane (P>0.05). Later in ovarian the staining intensity of MMP-2,9is stronger than in purple blue 103,500 stoves, and relating with theactivity of the ectopic endomemebrane (P<0.05) oMMP-2,MMP-9 and their activity can be sensitively detected by Gelatinzymogra_rn analysis in sera and ascites. MMP--2, 9 are highly expressedin patients than in control group(P<0.05) . The level of MMP--2 is higherin EMIII-IV and is nearly two times than in control (P <0.01). Theactivity of MMP-2 is higher than MMP-9 in sera of EM.The expression ofMMP-9 is low in control group. There is no statical difference amonggroups(P>0.05). MMP-9 increaces in abdominal cavity fluids of patients inEM III-IV.The expression of MMP-2,9 rise from rAFS I to IV.The results of RT-PCR are : The mR_NA of TMP-I,2 are expressed inall specimens,but the mRNA of MMP-2,9 in are only expressed in somespecimens. The expression and intensity of MMP-2,9mRNA in ectopicmembrane are equal to normal membrane,and there are no statical difference(P>0.05) .The expression of MMP--2,9 are higher than in controlgroup(P<0.05)o TIMP-1,2mRNA can be tested in endomembrane and tissues,whichare higher in ectopic membrane than in normal membranee (P<0.05). Theexpression of TIMP-lmRNA is slightly lower than the control group,but there isno statistical difference (P>0.05).The results of animal experiments showed : First, the transplanted tissueswere larger and cystic in animal models,full of yellow fluids.We can find uterineepithelium,glands and interstitial ,which told us the models of transplantationwere sucessful. Second,the size of tissues in pyrogallic acid groupandNafarelin Group were atrophied after drug therapy ,and there was staticaldifferences among groups. (P<0.05)o Third, in pyrogallic acid group,therewere decresed 腺腔 glands and altered fomation in light microscope .While inelectron microscope,there was changes of cellconection, orgenelle andappearance of atopsis body.There was a stronger role in C, D groups than inwith A, B Group (P<0.05) among diffenrant doses groups. Fourth, MMP-2,9and TIMP-1,2 in membrane were tested :they were expressed in normal rats tosustain the growth and dropping of membrane. There were a higherexpression of MMP-2,9 and a lower expression of TIMP-1,2 in controlgroup,showed that normal rats express TIMP-1,2 ,which had a statical differentwith group of sham operation (P<0.05).They enhance the ability oftransplantatin and come into ectopic region. Fifth,there were lower MMP-2,9and higher TIMP-1,2 in drug group.It made endomembrane atropy. Sixth,pyrogallic acid and Nafarelin can reduce ability of FSH, LH. Pyrogallic acidcan make FSH, LH reduce to normal level .E2 was slightly lower inpyrogallic acid than in control group, and there was no statical difference(P>0.05).E2 was significantly decreased in Nafarelin group,and there was astatical difference (P<0.05).My studies show that the excessive expression of MMP-2,9 in ectopicendomembrane makes it more invisive .But the expression of TIMP-1,2decreses,which leads to higher MMP/TIMP.Higher MMP/TIMP is related to thegrowth of membrane from both menstruaiton and fallen from uterus. MMP/TIMPis critical in the pathogenesis and development of the disease. MMP-2,9are better markers of EM and may be a sensitive indicator of EM.Pyrogallic acid can decreas the expression of MMP-1 and increase theexpression of TIMP-I.lt thereby reduces the ability of degradation ofexternal matrix by ectopic membrane.Inducing TIMP-1,2 or blockingMMP-2,9 artificially can hold the development of EM, which may become anew way of treatment for EM.