Studies Of Expression And Regulation Of Heparin-binding Epidermal Growth Factor(HB-EGF)in Human Endometrium,Early Villi And Decidua

Objective: (1) To investigate the expression and localization of heparin- binding epidermal growth factor (I-LB-EGF) in human normal endometrium during menstrual cycle and the first-trimester villi trophoblast and decidua, and evaluate its significance in endometrial growth, the implantation of the human blastocyst and early placental growth and development. (2) To study the expression of FIB- EGF in human uterine stromal cells in vitro culture, and its differential regulation by estradiol and progesterone. (3) To investigate the expression of epidermal growth factor receptor (EGFR) in human preimplantation embryos and to explore the effect of EGFR on human embryos growth, development and implantation. Materials and Methods: (1) 56 samples of endometrium of fertile women and 18 trophoblast cells and decidua at about seven to eight weeks gestation were detected. The expression and localization of HIB-EGF was examined by immunohistochemical method (LSAB) in human endometrium during the proliferative and secretory stage, and the first trimester chorionic villi and decidua. (2) Eight samples of endometrium of fertile women in the proliferative phase were detected. Stromal and epithelial cells were separated by enzymatic digestion method. The stromal cells were cultured in vitro, and passaged for secondary culture, and passaged to Corning culture dishes to grow for 24 hours. Then E2 and -4- P were added to the on罡tured system for 24,48 hours and In differentconcentrations of】p壬二三二(17B－estradiol,10',10'10"'mol几) P ( 10''10－''10－'mol／L),The variations of＊theions luorescence on the cells were detected with detectorof the laser scanning confocal microscope(LSCM)．(3)To detect the localizationand expression ofE哑R In human unfertlllzed oocytes and prelmplantatlonembryos with Indirect lmmunofluorescence assays(IIF)． Results:(l)m－EGF canbedetectedlnthe human endometrlum In botheplthellal and stromal cells,aM Inthe chorlonlc vllll trophoblast cells and decldua,m－EGF protein located on cell membrance and Inthe cytoplasm,and expressionof册－EGF Inhuman endometrlumhad periodical change duringthe menstrualcycle．The staining of皿－EGF protein was mainly seen In secretory phaseendometrlum with its peak Inthe lmplantatlon window stage and In throphoblastssfld d6Cldlls Of68fly pf6gflsllCX Stsllllflg fof HBHB－EGF WSS CSpCC13lly IOC8llZ6d tothe aplcal surface of＊thee lumlnal and glandular eplthellum during the［mplantatlonwindow stage．(2)E。or P significantly promoted the expression of HB－EGF Inhuman endometrlal stromal cells,at the concentrations of 10'or 10－'mol／L,andthe luorescence Intensity Increased after E。or P treatment and reached thenaxlsm 48 hours later,the e旺ct ofP onthe expression ofHBHB－EGF was more thanit ofE；(P＜001)．(3)There was EGFR expression In human unfertll；zed oocytesand prelmplantatlon embryos(from 2－cell to 10－cell stages)．EGFR protein locatedon the cell membrance surface,and there was no obvious variation In Intensity ofstaining Conclusions:(l)It Is suggested that HB－EGF was synthslzed and secreted Inhuman endometrlum and placenta,andm－EGF seems to play a regulative role Inthe Induction ofhuman endometrlal proliferation and dlfferentlatlon and have animportant function Inthe lmplantatlon ofthehuman blastocyst and early placentalgrowth and development．(2)It was Indicated that the。pression of HB－EGF inthC CfldoffiCtfl31 StfoffiBI CCllS WCfC pfoffiotCd by EZ 3fld P lfl VltfO．ThC CXpfCSSIOfl －5－ of HB-EGF may play an important role in the proliferation and differentiation of th...