The Molecular Study Of The Expression Of Vascular Endothelial Growth Factor(VEGF) And Matrix Metalloproteinases(MMPs) In The Endometrium Of Patients With Perimenopausal Anovulatory Dysfunctional Uterine Bleeding(DUB)

ObjectivesTo investigate the effects of estrogen and progesterone on the expression of ER,PR.MVD,vascular endothelial growth factor(VEGF) ,Matrix Metalloproteinases-9 (MMP-9) and tissue inhibitors of matrix metalloproteinases-1 (TIMP-1)in the endometrium of ovariectomized rat. To investigate the expression of estrogen receptor (ER),progesterone receptor (PR),Collagen IV and matrix metalloproteinase (MMPs)in the endometrium of perimenopausal patients with anovulatory dysfunctional uterine bleeding (DUB) . To evaluate the roles of cytokine , sex hormone and matrix metalloproteinase-9 in endometrial stromal cells of dysfunctional uterine bleeding by VEGF antibody and progesterone.-Methods1.The ovariectomized rat were injected intramuscularly with E ,P, E+P and oil respectively, the expression of ER ,PR ,CD34, VEGF, MMP-9 and TIMP-1 in the endometrium were detected with immunohistochemistry ,RT-PCR and/or Western blot. 2.Immunochemical SP technique was used to determine the expression of ER, PR, collage IV, MMP-9 and TIMP-1 in the endometrium of patients from 20 cases with normal menstrual cycle and 66 cases with anovulatory DUB.3. Concentrations of MMP-9 were determined by ELISA. Sisteen patients with DUB were included as study group and eleven patients without DUB as control group.Results1. Compared with the no-operation group, the expression of ER,PR,CD34, VEGF, MMP-9 are decreased in C,P and oil groups whereas elevated in E and E+P groups.In the HRT group ,the level of ER,PR,VEGFmRNA 及 MMP-9mRNA in the endometrium are positively correlated with estrogen treatment, whereas MMP-9level may decrease if treated with progesteron on this basement.2. Expression of ER, PR is significantly higher in the endometrium with simplex and complex hyperplasia in comparison to the normal proliferative endometrium (P<0.05),MMP-9 immunoreactivity in glandular epithelial cells and MMP-9/ TIMP-1 ratio is significantiy higher in endometrium with simplex and complex hyperplasia in comparison with the proliferate endometrium (P<0.01), expression of collagen IV is significantly lower in the endometrial stroma of DUB patients. MMP-9 is positively correlated with ER (r=0.605,P<0.05) and PR (r=0.697,P<0.05).3.Levels of MMP-9 in the DUB group was significantly higher than the control group. Secretion of MMP-9 in each group was decreased in culture with progesterone and VEGF antibody, level of MMP-9 in DUB group was significactly decreased than the control group in the higher concentration of progesterone secretion. Levels of MMP-9 in each group decreased in culture with VEGF antibody, but the level of MMP-9 in each group has no difference between 24h,48h after cultured with VEGFantibody. Conclusions1. The results suggested that the physiological and pathological changes in the endometrium are under both the effects of ovarian sex hormones and the related molecules in the microenvironment through auto-endocrine and para-endocrine mechanisms. 2. The likely causes of abnormal bleeding in DUB patients with simple and complex endometrium hyperplasia are associated with abnormal expression of endometrium glandular ER and PR under the action of sexual hormone and with upregulation of activity of MMP-9 in the endometrium which cause over degradation of the ECM.3. VEGF may play a role in pathogenesis of DUB by increasing the secreation of MMP-9 in the endometrial stromal cells, progesterone can decrease the secretion of MMP-9 and it may restrain the development of DUB.