Effects Of Endometriosis Peritoneal Fluid Of Different Clinical Stages In Maturation Of Cord Blood Monocyte Differentiate Into Dendritic Cells

Background:Endometriosis (endometripsis, EMs) is a growing function of endometrium (glands and stroma) grows outside the uterine cavity, infiltrate, proliferate, diffuse, recurrence, cause pains and infertility. EMs is a common disease of childbearing age women.The incidence of this disease has increased considerably in the past few years,even in Adolescent girls and postmenopausal women.1860, The researchers were firstly found EMs,so far,but the pathogenesis is also unclear, resulting in difficulties of treating this disease. Therefore, Researching of the pathogenesis is necessary. At present, Sampson's theory is still the leading theory of endometriosis,but recent studies suggest that menstruation reflux is the incentive of endometriosis, the pathogenesis of endometriosis is relative with immune abnormalities, even EMs are autoimmune diseases.Dendritic cells (dendritic cell, DC) is the most powerful antigen-presenting cell(APC), involved in antigen recognition, processing, and delivery,and in the immune response induced in an impotant position. DC has a dual function, it induce immune response and also induce immune tolerance. Immunologic function of DC and maturation are relational, in DC development,it can be divided into the Immature DC and mature DC. Immature DC can induce the formation of CD4+ T cells, secreted immunosuppressive cytokines IL-10; mature DC induced T cell proliferation, promote cell mediated mediated immune response. DC participate immunization, inflammation, tumor immunity, graft rejection, emergency response and other physiological and pathological processes. When the quantity of DC reduced or the imbalance between immature DC and mature DC, it will lead to a series of immune-related diseases, such as:asthma, tissue disease, colitis, cancer, etc., and has attracted people's attention. In recent years, the role of DC in EMs catches more and more attention in immune abnormalities research. This study is based on the theory that mononuclear cells extracted from umbilical cord blood could differentiate into dendritic cells under certain conditions (GM-CSF+IL-4). And the main content of this study is to discuss how peritoneal fluid of EMs(ePF) effects mononuclear cells differentiate into DC.Objective: 山西医科大学硕士学位论文This study will study the following aspects after mononuclear cells be intervented to differentiate into DC by ePF;will investigate the presence of IL-6,IL-10 in peritoneal fluid and determined if differences in the concentration of these cytokines existed between women with and without endometrisis. and explore the roles of these cytokines in different clinical stage of endometriosis.Method:1.30 case of EMs visited the Obstetrics and Gynecology Department of Second Shanxi Medical University were chosen as research objects, and 15 cases of Uterine Myoma, and then all of peritoneal fluid as research objects were collected. The status of all patients since July, 2008 to December,2009.2. Collected from January,2009 to April,2009 of collected umbilical cord blood from neonatus with Second Hospital of Shanxi Medical University between the normal obstetric delivery.3. Separation of mononuclear cells cultured for 7 days in the culture medium by adding cytokines GM-CSF+IL-4 and peritoneal fluid (EMs peritoneal fluid and peritoneal fluid of the control group); flow cytometry examined proportion of cells that express CD la and CD83. ELSIA detected IL-6 and IL-10 levels of peritoneal fluid and DC supernatant.Results:1. Different clinical stages of peritoneal fluid of endometriosis have different effects in process of cord blood monocyte differentiate into Dendritic Cells.1.1 After cultured 24h, the dish appeart scattered cells polymer, cell type was smaller and round, the main cells were adherent, only a small number of cells were semi-suspension, after 3-5 days in the cPF Group,the suspended cells increased with irregular shape, cell volume increased, aggregate-like clusters, with the culture time extended,the number of suspended cells increased.1.2 The seventh day, cells become larger, there are protuberance on the cell surface; but in ePF group,most cells were adherent, non-suspension and did not aggregate clusters.Adherent cells were mainly macrophages shape which is round or oval nucleus, and cytoplasm is rich, within some cell contain particles or vacuoles, which are more evident between stageⅢ-ⅣEMs and stageⅠ-ⅡEMs.1.3 In freshly isolated monocytes,proportion cells that express CD1a and CD83 were low; At 3rd day and 7th day, proportion of CD1a cells increased, the proportion of CD1a cells in stageⅠ-ⅡEMs lower than stageⅢ-ⅣEMs and cPF group's, the difference was significant (P <0.05),Ⅲ-ⅣEMs and cPF group's are compared, there was no significant difference (P> 0.05).at 3rd day and 7th day,within group compared, the proportion of CD83 cells increased, the difference was significant (P<0.05), while compared among groups, stageⅠ-ⅡEMs more increased than stageⅢ-ⅣEMs and cPF group's, the difference was significant (P<0.05), stageⅠ-ⅡEMs low cPF group,there was no significant difference (P> 0.05).2. the relationship between the levels of IL-10 level and clinical stage of endometriosis2.1 In ePF group, in peritoneal fluid,in levels of IL-10,stageⅠ-ⅡEMs are higher than cPF group, the difference is significant (P<0.05). stageⅠ-ⅡEMs are higher thanⅢ-ⅣEMs, there is no significant difference (P> 0.05).2.2 In the supernatant of 7th cultured days, in levels of IL-10, stageⅠ-ⅡEMs are higher, the difference is significant (P<0.05).Ⅲ-ⅣEMs has no significant change (P>0.05). cPF group are higher, the difference is significant (P<0.05). stageⅢⅣEMs,EMs are lower than cPF group, there is no significant difference (P> 0.05).3. the relationship between the levels of IL-6 level and clinical stage of endometriosis3.1 In ePF group, in peritoneal fluid,in levels of IL-6,stageⅠ-ⅡEMs andⅢ-ⅣEMs are higher than cPF group, the difference is significant (P<0.05).3.2 In the supernatant of 7th cultured days, in levels of IL-6, all groups are higher.Ⅰ-ⅡEMs has no significant change (P>0.05).Ⅲ-ⅣEMs are higher, the difference is significant (P<0.05).cPF group has no significant change (P>0.05). stageⅢ-ⅣEMs,EMs are lower than cPF group, there is no significant difference (P> 0.05).Ⅰ-ⅡEMs andⅢ-ⅣEMs are higher than cPF group, the difference is significant (P<0.05). stageⅢ-ⅣEMs are higher thanⅠ-ⅡEMs, the difference was significant (P<0.05).Conclusion:1. the mumbers of DC in EMs groups are lower than in the control group; groupⅠ-ⅡEMs are non-mature DC type, and groupⅢ-ⅣEMs are mature DC type.2. in the peritoneal fluid interferenced,groupⅠ-ⅡEMs are higher in IL-10 level,clued that immune state is immne suppress in ealy stage of EMs.3. groupⅢ-ⅣEMs are higher in IL-6 level, clued that immune state is immunologic activation in later period of EMs.