| Objective:To detect the Gremlinl mRNA level in granulose cells in women wish diminished ovarian reserve (DOR), and analyze the relationship between the Gremlinl (GREM1) mRNA level and in vitro fertilization-embryo transfer (IVF-ET) outcomes. We also investigated the role of GREM1in cumulus cell expansion.Methods:22women with diminished ovarian reserve and22women with normal ovarian reserve (NOR) were examined. We measured the gene expression of GREM1and cumulus expansion-related transcripts PTX3, HAS2, PTGS2, and TNFAIP6in granulosa cells from these44infertile women under38years old. The doses of gonadotropin used, the number of follicles and eggs, the fertilization rate and the rate of high quality embryos were recorded, and the pregnancy rate was followed up. In vitro, the KGN cells strains (n=3) were treated with follicle-stimulating hormone (FSH) and recombinant gremlin for0,4,6and24h to evaluate the effects of exogenous gremlin on the mRNA expression of PTX3, HAS2, PTGS2and TNFAIP6. We next used a small molecule inhibitor to explore the involvement of TGF-β signaling pathway in gremlin-mediated cumulus expansion-related transcripts induction. SB431542is a inhibitor of SMAD2/3signaling pathway. We pretreated KGN cell strains (n=3) for1h with or without SB431542followed by treatment with FSH and recombinant gremlin for6h to detect the effect of SB431542on the expression of PTX3, HAS2, PTGS2and TNFAIP6.Result:GREMl, PTX3, HAS2, PTGS2, and TNFAIP6were down-regulated in women with DOR versus NOR (p<0.01). In vitro, Gremlin significantly (p<0.01) induced PTX3, HAS2, PTGS2and TNFAIP6mRNA expression, although the time course of induction varied for each gene. By six hours, gremlin significantly induced all genes. And SB431542blocked gremlin-mediated HAS2induction in all KGN cell strains tested (p<0.01).Conclusions:Decreased GREM1expression in granulose cell has an underlying defect in cumulus cell expansion, and resulted in diminished ovarian reserve. |
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