| Embryo is production of the two histocompatibility individual mating,which is expresses antigens foreign to mother,so has been viewed as allograft.Immunological rejection is a basic reflection to the allograft from the body's immune system.In the process of normal pregnancy,embryos can implant,grow in the matrix,and acquire immune nutritional and immune protection from the maternal immune system.The maternal-fetal immune tolerance is the exception in immunology.When the maternal-fetal immune tolerance is destroyed,the immune rejection can be cause and lead to pregnancy loss.So the maternal-fetal immune tolerance is essential for successful pregnancy.The characteristics of the maternal-fetal interface was the premise of the maternal-fetal immune tolerance.The decidua of maternal-fetal interface contains many kinds of inflammatory cell,which has the potential that triggered an immune response against pathogens.Early pregnancy fetal extra villous trophoblasts invade the decidual tissues,direct contact with maternal decidual cells,forming a subtle maternal-fetal immune microenvironment.Decidual immune cell(DIC)is important for the immune activity of cells,Including the human T lymphocyte,uterine natural killer cell(u NK cells),Macrophage(M?),etc.Maternal cells,fetal cells and their secretion of cytokines,growth factors,hormones,such as micro environment constitute the maternal-fetal interface.As a group of immune cells of the maternal-fetal interface,DICs play an important role in the maternal-fetal immune balance.Many studies have found that Th17 cells of peripheral blood and decidua group was obviously higher in the patients with recurrent spontaneous Abortion(RSA)than that of normal pregnancy group,and with the rise of IL-17 and IL-23.Compared with normal pregnancy group,our previous study also found that IL-23,IL-6 IL-17 cytokines increased significantly in peripheral blood and decidua tissues of recurrent spontaneous Abortion.IL-23 / IL17 axis is a classic inflammation axis,IL – 23 activate JAK-STAT signal pathway through combined with IL-23 R,signal transduction and transcriptional activation factor 3 phosphorylation and transferred to the nucleus,Raised the ROR gamma t transcription,which is Th17 cells specific transcription factor,and further increase the proliferation and differentiation of Th17 and increase IL17 secretion.Now we propose to investigate whether IL-23 can activate the STAT3 in human pregnancy maternal-fetal interface,and increase the secretion of IL-17 factors and enhance inflammation to cause the failure of pregnancy,which to clarify the regulation mechanism of IL-23 in decidua immune cells.As a kind of adrenal cortical hormone,prednisone is widely applied in some autoimmune diseases and the treatment of recurrent spontaneous abortion.Treg/Th17 balance is very important for maternal immune tolerance of fetus.Treg/Th17 balance is exist in maternal-fetal interface.Normal pregnancy is given priority to with Treg migration,and Th17 cells increased in recurrent spontaneous abortion patients.Whether prednisone can regulate Treg/Th17balance? We explore the role of prednisone in Treg/Th17 balance by prednisone drug-containing serum on spleen lymphocytes.Part One The characteristic of phosphorylation of STAT3 and cytokine changes in the maternal-fetal interface of recurrent spontaneous abortionObjective: Comparison of the maternal-fetal interface level of STAT3 protein phosphorylation,IL-17 receptors and the change of related cytokines between spontaneous abortion patients and normal pregnancy women.Method:Collect the peripheral blood and decidual tissues of recurrent spontaneous abortion and normal pregnancy,to detect and compare the decidual tissues STAT3 phosphorylation and protein expression in two groups through Western Blot(Wb)and Immunohistochmeistry(IHC).To detect the levels of IL-6,IL10,IL-17,and IL-23 related cytokines in peripheral blood and decidua tissues by Enzyme linked immunosorbent assay(Elisa).Result: The protein levels of STAT3 protein phosphorylation and IL-17 R in maternal-fetal interface decidual tissue of recurrent spontaneous abortion are higher than in normal pregnancy group.The levels of IL-10 are lower than normal group in abortion group,and the levels of IL-17 IL-23 in abortion group are higher than normal group.The level of IL-23 and p-STAT3 were positively correlated,the level of IL-17 has a consistent trend with the levels of IL-23 and p-STAT3.IL-23 increased may cause STAT3 activation and enhanced the IL-17 levels in abortion group.Conclusion: IL-23 May strengthen the inflammatory response by promoting STAT3 phosphorylation,which is involved in the occurrence of recurrent spontaneous abortion.Part Two The effect of IL-23 on the Treg/Th17 balance in the decidua immune cells of maternal-fetal interfaceObjective:To discuss the effect of IL-23 protein and IL-23 antibody on human pregnancy maternal-fetal interface decidual immune cells in the local microenvironment.Method: Preparation human pregnancy decidua immune cells through the primary cell culture method.Using the anti-CD3 antibodies and anti-CD28 to activate the original generation decidual immune cells.The decidual immune cells were processed by anti-IL-23 antibody and IL-23 protein respectively for 24 hours.Setting up blank control group,IL-23 protein processing group,anti-IL-23 antibody treatment group respectively.Flow cytometry analysis the levels of Treg and Th17 among the three groups.The expression levels of STAT3,p-STAT3,IL-23 R were detected by Western Blot.After cell culture for 12,24,and 36 hours,IL-1??IL-4?IL-10 and IL-17 levels of the culture supernatant were detected by Elisa,explore the change of the related cytokines in cell culture with the time.Result: IL-23 protein promote Treg/Th17 balance to the offset in the direction of Th17 in decidual immune cells of maternal-fetal interface.As a result,raise the expression of STAT3 phosphorylation and IL-23 receptor,promote the secretion of proinflammatory factor such as IL – 1? and IL-17.On the contrary,Anti-IL23 antibodies promote pregnancy maternal-fetal interface decidual Treg edge in immune cell differentiation,inhibit Th17 cell differentiation.Reduce the activation of STAT3 protein and the expression of IL-23 receptor proteins,promote the secretion of inflammation suppression factor such as IL – 4 and IL-10.Conclusion: Anti-IL-23 antibodies make for Th17 / Treg balance offset in the direction of Treg cells in decidual immune cells of pregnancy maternalfetal interface,Promote inflammation suppression factor secretion,reduce the activation of STAT3 protein and protein expression of IL – 23 R.Part Three The effect of Prednisone drug-containing serum of rats on the spleen lymphocytes of rats in Treg/Th17 balance and the related transcription factors Foxp3,ROR?tObjective: Explore the effect of prednisone drug-containing serum on spleen lymphocytes in Treg/Th17 balance and its relevant transcription factor Foxp3,ROR?t.Method: Preparation of rat blank and prednisone drug-containing serum,with high performance liquid chromatography(HPLC)method to detect the drug-containing serum of prednisone drug active ingredient.Isolated spleen lymphocytes in rats by lymphocyte separation medium method.Using the rat blank and prednisone drug-containing serum training rats spleen lymphocytes Separately,the spleen lymphocytes were divided into four groups According to the join order of cytokines and whether they contain prednisone drug-containing serum.Respectively is group a: blank serum culture after 24 hours and add to IL-6,IL-23,TGF – ? to the cells,collecting cells after culture 48 hours;group b:prednisone drug-containing serum culture after 24 hours and add to IL-6,IL-23,TGF – ? to the cells,collecting cells after culture 48 hours;group c: Joined the IL-6,IL-23,TGF-? cell factors to the blank serum to training rats spleen lymphocytes for 48 hours,and then in culture medium containing blank serum cultured cells and collected cells after 24 hours;group d: Joined the IL-6,IL-23,TGF-? cell factors to the blank serum to training rats spleen lymphocytes for 48 hours,and then in culture medium containing prednisone drug-containing serum serum cultured cells and collected cells after 24 hours.Detect the Treg/Th17 balance of each group cells and the related gene Foxp3,ROR?t transcription level.Result: The result comparison show that The proportion of Treg cells of the spleen lymphocyteswere increased the prednisone drug-containing serum groups,and Th17 cells decreased.On the other hand,The proportion of Th17 cells rised and Treg cells decreased in blank serum group.With prednisone drug-containing serum cultivation to spleen lymphocytes in advance can effectively inhibit cytokines in lymphocyte CD4 + T cells to Th17 direction of entrainment.Conclusion: Prednisone can affect Treg/Th17 balance to be offset in the direction of Treg in spleen lymphocytes. |
PDF Full Text Request