The Effects Of Gonadal Hormone On IL-18Secretion And Expression Of Goat Endometrial Cells

Endometrium similar to the pneogaster and gastrointestinal tract mucous membraneconstitute the first line of defense of body, it is an important component of the body's mucousmembrane immune. Gonadal hormone has the important regulating function on embryoimplantation and establish pregnancy and endometrial cells' growth and secretory activity.Interleukin-18(IL-18) is a pleiotropic and proinflammatory cytokines that new discovery inrecent years and express in the endometrial cells and mononuclear cell and macrophage inhumans and mice. It has proved that IL-18is a paracrine cytokines, IL-18play a significantroles in regulating physiological function of endometrial cells and regulating immunetolerance in maternal-embryo interface and maintain of normal pregnancy and the process ofembryo implantation, while the research reports about the regulating function of gonadalhormone on IL-18secretion and expression of endometrial cells is less.In order to research IL-18secretion and expression activity of goat endometrial epithelialcells (EEC) under the action of gonadal hormone and regulating action of endometrial stromalcells (ESC) on IL-18secretion and expression activity of EEC, this test based on the nest dishculture method and make use of immortal goat EEC and ESC to construct cells complex layermodel in vitro, the aim is to detect and analyse regulating action of E2and/or P₄on IL-18secretion activity in EEC and the effects of ESC on it through ELISA, Western blot,semiquantitative RT-PCR and immunochemistry, the results as following:1. Detection of ELISA show: Compared with no gonadal hormone treatment blank controlgroup, independent treatment of P₄or together treatment with E2will extremly significantenhance the IL-18secretion activity (P<0.01), while E2alone treat has no significantdifference; when EEC co-cultured with ESC in the nest dish culture system, independenttreatment of E2or together treatment with P₄can significant inhibit IL-18secretion activity inculture supernatants of EEC (P<0.05), while P₄alone treat has no significant difference. IL-18secretion level of EEC co-cultured with ESC significant higher than EEC cultured alone whenit take no gonadal hormone treatment, the detection of immunocytochemistry obtain similar results. The detection of Western blot indicated that IL-18secreted by EEC main exist in theform of biological activities and its molecular weight is18.3kDa.2. Semi-quantitative RT-PCR research results show: Independent treatment of E2cansignificant enhance IL-18mRNA expression level in EEC cultured alone (P<0.05),independent treatment of P₄or together treatment with E2will further enhance the IL-18mRNA expression level (P<0.01); when EEC co-cultured with culture supernatants of ESC,independent treatment of P₄or together treatment with E2can extremly significant inhibitIL-18mRNA expression level of EEC (P<0.01), while E2treat alone has no significantdifference of IL-18mRNA expression level. When it take no gonadal hormone treatment,IL-18mRNA expression level of EEC co-cultured with ESC significant higher than EECcultured alone, this indicated that gonadal hormone and ESC regulate IL-18expression levelat the transcriptional level.