The Role Of Histidine Triad Nucleotide Binding Protein 1 In Endometrial Stromal Cell Decidualization In Mice

Objective: Embryo implantation is initiated following the adhesion of blastocysts to uterine epithelial cells.Successful embryo implantation depends on the interaction between a activated blastocyst and a recipient uterus.After implantation,the endometrial stromal cells surrounding the embryos are initiated,spontaneously proliferating and differentiating into decidual cells.According to statistics,about 20% of failed pregnancies are caused by abnormal decidualization during embryo implantation.Therefore,the normal progression of decidualization is critical for the establishment and maintenance of pregnancy.Histidine triad nucleotide binding protein 1(Hint1)has been found to play an important role in the regulation of gene transcription pathways and metabolism.To explore the regulatory role of Hint1 in endometrial stromal cell decidualization in mice.We collected decidual tissues from normal pregnancy,pseudopregnancy and artificially induced deciduallization in KM mice.Immunohistochemistry,western blot,in situ hybridization,and RT-qPCR were used to investigate the expression pattern of Hint1 in early pregnancy in mice.Afterwards,mouse endometrial stromal cells wereisolated and cultured.SiRNA targeted to Hint1 was use to knock down expression of Hint.Western blot,RT-qPCR,and liquid Chromatograph Mass Spectrometer(LC-MS)methods were used to investigate the regulation and mechanism of Hint1 on decidualization of endometrial stromal cells.Methods:(1)The Immunohistochemistry,in situ hybridization,western blot,and RT-qPCR were used to investigate the Hint1 expression in early pregnancy,establishment of the pseudopregnancy and artificially-induced decidualization mouse model.(2)si-Hint1 was used to knockdown Hint1 expression in decidualized stromal cells in vitro and the influence of Hint1 exoression on during the endometrial stromal cells decidualization was analyzed.(3)LC-MS was used to analyze the effect of Hint1 on stromal cell lipid metabolism.Results:(1)The expression of Hint1 gradually increased during the progress of early pregnancy from day 1 to day 8,and the expression at the implantation sites was significantly higher than the inter implantation sites.(2)The expression of Hint1 was significantly higher in the endometrium of artificially-induced decidualization mice than the control.(3)The interference with Hint1 by siRNA significantly influencedthe decidualization and lipid metabolism in mouse stromal cells in vitro.Conclusion: Hint1 is spatio-temporally expressed during cell decidualization and decidualization may be associated with lipid metabolism.