The Study On The Effect Of Macrophage And Epithelial-mesenchymal Transition Induced By It On The Pathogenesis Of Adenomyosis

Background:Adenomyosis is a common disease which is benign and non-neoplastic for reproductive or perimenopausal women.The biological behavior of adenomyosis is similar to malignant tumor.In recent years,the study show that epithelial-mesenchymal transition(EMT)induced by M2 macrophages plays important role in tumor progression.Previous study demonstrate that macrophages infiltrate in adenomyosis lesions,and some studies show that EMT progress occur in adenomyosis lesions.Thus,we suspect that macrophage and EMT induced by it are important of adenomyosis process on the pathogenesis.Chapter 1 The relationship between macrophages and epithelial-mesenchymal transition(EMT)of adenomyosis lesions is close.Objective:1.To validate the phenomenon of epithelial-mesenchymal transition(EMT)in adenomyosis lesions.2.To detect the distribution of macrophages in adenomyosis lesions.3.To observe the relationship between the phenomenon of epithelial-mesenchymal transition(EMT)and the distribution of macrophages in adenomyosis lesions.Materials and methods:Stain the clinic specimen of normal endometrium,eutopic and ectopic endometrium of adenomyosis by immunohistochemistry and immunofluorescence to detect the protein expression of E-cadherin,N-cadherin,CK7,vimentin,S100A4 and CD68.Then,analyse expression profile to find the proportion of epithelial-mesenchymal transition(EMT)in adenomyosis,simultaneously,observe the relationship between the phenomenon of epithelial-mesenchymal transition(EMT)and the distribution of macrophages in adenomyosis lesions.Results:1.The phenomenon of epithelial-mesenchymal transition(EMT)occurs in adenomyosis lesions,and the proportion of EMT is high.2.Mass of macrophages infiltrate in the adenomyosis lesions,and the distribution of macrophages is independent of the menstrual cycle.3.The distribution of macrophages is negative to correlate with the expression of epithelial makers,but positive to correlate with mesenchyme makers.Conclusion:The relationship between the infiltration of macrophages and the EMT of adenomyosis lesions is close.And macrophages are important participants in EMT process of adenomyosis lesions.Chapter 2 Macrophages induce epithelial cells of endometria to undergo epithelial-mesenchymal transition(EMT).Objective:1.To detect whether or not macrophages can induce Ishikawa cells to undergo EMT.2.To detect whether or not macrophages can induce eutopic endometrium epithelial cells of adenomyosis or normal endometrium epithelial cells to undergo EMT.Materials and methods:Co-culture macrophages with Ishikawa cells,eutopic endometrium epithelial cells and normal endometrium epithelial cells.Detect the protein expression of E-cadherin,N-cadherin,CK7,vimentin,S100A4 by Western Blot in Ishikawa cells,eutopic endometrium epithelial cells and normal endometrium epithelial cellsbefore and after co-culturing.Contrast the invasive ability of Ishikawa cells using cell matrigel invasion assay before and after undergoing EMT.Observe the changes of cellular morphology and cell junction of Ishikawa cells by ordinary optics microscope and transmission electron microscope(TEM)before and after co-culture.Detect the changes of proliferation and apoptosis of Ishikawa cells by FCM.At last,find the changes of signaling pathway of Ishikawa cells after co-culture with macrophages undergoing EMT.Results:1.Macrophages induce Ishikawa cells to undergo EMT.2.Macrophages induce eutopic endometrium epithelial cells and normal endometrium epithelial cells to undergo EMT.3.The potential ofEMT is more intense in eutopic endometrium epithelial cells than normal endometrium epithelial cells after co-culture with macrophages.Conclusion:Macrophages can induce endometrium epithelial cells to undergo EMT.And,comparing with normal endometrium epithelial cells,the potential of EMT process is more intense in eutopic endometrium epithelial cells after co-culture with macrophages.Chapter 3 Endometrium epithelial cells induce macrophages to polarize to M2 in the co-culturing systemObjective:1.To detect whether or not Ishikawa cells induce macrophages to polarize to M2.2.To detect whether or not eutopic endometrium epithelial cells and normal endometrium epithelial cells induce macrophages to polarize to M2.Materials and methods:Detect the changes of molecular and genetic expression of CD 163,IL-10 and MMP12 which are markers for M2 by FCM and polymerase chain reaction(PCR)in macrophages after co-culture with Ishikawa cells,eutopic endometrium epithelial cells and normal endometrium epithelial cells.Results:1.Ishikawa cells induce macrophages to polarize to M2.2.Both eutopic endometrium epithelial cells and normal endometrium epithelial cells can induce macrophages to polarize to M2.3.A further polarization to M2 of macrophages was induced by normal endometrium epithelial cells,comparing with eutopic endometrium epithelial cells.Conclusion:Ishikawa cells,eutopic endometrium epithelial cells and normal endometrium epithelial cells all can induce macrophages to polarize to M2.Simultaneously,a further polarization to M2 of macrophages was induced by normal endometrium epithelial cells,comparing with eutopic endometrium epithelial cells.