The Study On Invasion Property Of Eutopic Endometrium And Immunology In Endometriosis Pathogenesis

The study on invasion property of eutopic endometrium from women with endometriosis and the immunology in the endometriosis pathogenesisBackgroundEndometriosis is a common gynecologic disorder of the reproductive age women by the presence of endometrial tissue outside the uterine cavity. Although no single theory can explain all cases of endometriosis, the most commonly accepted theory is Sampson's theory of retrograde menstruation. But the exact mechanisms of the initial interaction between refluxed endometrium and peritoneum are still not completely understood. Several in vivo and in vitro models have been developed to study these interactions between endometrium and peritoneum. Some researches proposed baboon or nude mice as in vivo culture model for the study of endometrial-peritoneal implantation. And others used amnion membranes or biopsies of human peritoneum as in vitro model to study the potential interaction, but the conclusions are controversial. In our laboratory, we have developed a novel model of co-culture of endometrial-peritoneum histological model, which have advantages of good imitation and are suitable for comparison of endometrium and peritoneum in patients and normal women. It also could be observed dynamically of the initial stage of pathogenesis of endometriosis such as adhesion, invasion and angiogenesis. Sampson' s theory can not explain the fact that retrograde menstruation occurs in 70%-90% of women, but the prevalence of endometriosis is much lower. Recent years, based on systemic research data about endometriosis, professor Lang Jinghe, the eminent scholar from PUMCH, proposed a new theory to endometriosis, that is the "Eutopic endometrium determination theory". The essential of the novel theory is that the eutopic endoemtrium from women with endometriosis has some fundamental differences compared to eutopic endometrium form women without endometriosis. This differences determine the susceptibility do develop endometriosis with potential of adhesion, invasion and angiogenesis. Immune theory is another theory to interpret the above-mentioned difficulty of Sampson's theory. Its content includes endometriosis is associated with changes in both cell-mediated and humoral immunity. This leads to inadequate removal of refluxed menstrual debris. Macrophages that would be expected to clear endometrial cells from the peritoneal cavity appear to enhance their proliferation by secreting growth factors and cytokines. We believe that these immunologic alterations are a consequence of the presence of endometrium in the peritoneal cavity that is to say it is the abnormal eutopic endometrium itself who changed the immunologic reaction.Objective1. To establish the endometrial-peritoneum histologic research model for endometriosis and observe the property of early invasion of endometrium to peritoneum and compare the difference of invasiveness between eutopic endometrium from women with endometriosis and that form women without endometriosis.2. To investigate the properties of eutopic endometrium which determine its potential of invasiveness from a perspective of molecular biology.3. To explore the molecular mechanism for endometrium from women with endometriosis to escape the immunologic surveillance and survive, implant and proliferate on peritoneum.Method1. Observe the invasion of endometrium into peritoneum after co-culture in vitro for different period of time under light microscope.2. Detect the level of mRNA or protein of MMP-3, MMP-9 in endometrium or culture supernatant using IHC and RT-PCR.3. Study the expression of mRNA and protein of ENDO-1 in endometrium and culture supernatant with IHC, RT-PCR and western blotting. ENDO-1 is believed to be able to change the function of macrophages and promote angiogenesis.4. Observe and analysis the proliferation and apoptosis of endometrium using IHC and TUNEL and the relationship with Fas/FasL, the apoptosis promoting system.5. Detect the secretion of sFasL in the supernatant of co-culture.Result1. After co-culture with peritoneum in vitro, the endometrium from women with endometriosis sustained and invaded more significantly than that from women without endometriosis.2. The levels of mRNA of MMP-3, MMP-9 in endometrium from women with endometriosis was more higher than that of women without endometriosis.3. Free MMP-3 protein could be detected in the co-culture supernatant.4. The expressions of ENDO-1 protein and mRNA in endometrim during secretory stage from women with endometriosis were higher than that from women without endometriosis.5. Endometrium from women with endometriosis during secretory stage was less sensitive to apoptosis. But the level of Fas mRNA has no difference between group of endometrium and control.6. In the supernatant of co-culture, more sFasL could be detected, which was believed to prompt the apoptosis of CTL and cause the immune tolerance of refluxed endometrium in peritoneal cavity.7. The proliferation activity of endometrium from women with endometriosis in secretory stage had no difference with that from women without endometriosis.Conclusion1. Endometrial-peritoneum co-culture model is suitable for study of early invasion of refluxed endometrium to peritoneum.2. The potential of endomtrium from women with or without endometriosis is different with the different expression of MMPs.3. The proliferation ability is of no difference between the endometrium from women with or without endometriosis, but the endometrium from women with endometriosis is less sensitive to apoptosis.4. In endometriosis group, more sFasL is secreted in supematant of co-culture than that in control group.