| Objectives:To explore the expression and localization of KiSS-1/GPR54system in theuterine endometrium, and its roles in the decidualization.Methods:1. To observe the expression and localization of KiSS-1and GPR54in mice uterusduring peri-implantation by immunohistochemistry, real-time quantitative PCRmethod.2. To observe whether the expressions of KiSS-1and GPR54are influenced byactivation embryo by using artificially induced decidualization model in vivo.3. To observe whether the expressions of KiSS-1and GPR54are regulated by thesteroid hormones by ovariectomized mouse and steroid hormone treatment.4. The expression levels of KiSS-1and GPR54were observed in mouse uterusduring artificial decidualization after endometrial stromal cells were isolated,cultured and induced by E2and P4in vitro.5. To observe the role of KiSS-1in the decidualization of stromal cells by RNAinterference technology.Results：1. Real-time quantitative PCR results showed that KiSS-1and GPR54mRNAexpression were lower during D1-D5of pregnancy. With the progress ofdecidualization (D6-D8of pregnancy), KiSS-1and GPR54mRNA expressionwere significantly increased compared with D1of pregnancy (P<0.01).Immunohistochemical results showed that KiSS-1and GPR54protein are mainlylocated in the luminal and glandular epithelium, and the levels of their expressionare low during D1-D4of pregnancy, which is consistent with the real-timequantitative PCR results. From the D5of pregnancy, the expression levels ofKiSS-1and GPR54protein were significantly increased, and mainly located inthe decidual tissues surrounding the embryo. 2. In the uterus of artificial decidualization, compared with the control group, theuterine horn injected with sesame oil gradually increases from day5to day8ofpseudopregnancy, but control group did not significantly change. In uterine tissueof artificially induced decidualization, The expression levels of KiSS-1andGPR54mRNA in the uterus of artificial decidualization gradually increased withthe progress of decidualization, a peak on day8of pseudopregnancy. In day5ofpseudopregnancy, KiSS-1and GPR54protein were mainly located in theluminal epithelium of uterus, and the signal is weak. KiSS-1and GPR54proteinsignificantly increased in the decidual tissue on day6of pseudopregnancy, andgradually increased with the progress of decidualization.3. The expression level of KiSS-1mRNA is relatively low in ovariectomized uterustissue, After E2, P4or E2+P4treatment, compared with the control group, KiSS-1mRNA expression was significantly increased (all P<0.01), a peak at3h afterinjection; Its expression did not significantly increase in the uterus treated withthe combination P4and its antagonist RU486(P>0.05); whereas further increasedin the uterus treated with E2and its antagonist Tam.4. Immunofluorescence assay showed that isolated stromal cells were expressedvimentin not cytokeratin protein. After the E2and P4combined treatment for24h-72h in vitro, stromal cells showed specific morphological characteristics ofdecidual cells; Western blot and quantitative PCR detection showed theexpression levels of Cyclin D3protein, PR protein and dPRP mRNA, which aredecidua marker molecules, significantly increased after48and72h culture invitro and KiSS-1and GPR54mRNA expression levels also gradually increaseswith the progress of decidualization.5. In the artificial decidualization model of uterine stromal cells in vitro, afterKiSS-1siRNA transfection, KiSS-1mRNA expression was significantlydecreased compared with the control group (scrambled group) in24h,48h,72hculture (P <0.01,0.05), which demonstrated that KiSS-1siRNA could specificallyinhibit the expression of KiSS-1mRNA. dPRP (mRNA), Cyclin D3(protein)and PR (protein) expression levels also significantly decreased after KiSS-1siRNA treatment, suggesting that down regulation of KiSS-1gene could significantly inhibit the decidualization of uterine stromal cells.Conclusion:KiSS-1/GPR54system was spatial-temporally expressed at the maternal-fetalinterface during early pregnancy, and their expressions were influenced by E2, P4anduterine decidualization; the decidualization of endometrial stromal cells significantlyinhibited after interference KISS-1gene, It suggested that KiSS-1/GPR54system wasinvolved in promoting uterine decidualization. |
PDF Full Text Request