Effect Of Hand2os1 In The Formation And Degeneration Of The Corpus Luteum In Mice

As a temporary endocrine organ,the corpus luteum plays an important role in regulating and maintaining pregnancy during the female reproductive cycle.It is formed by the rupture of the follicle after ovulation.Its main physiological function is to secrete steroid hormones,mainly progesterone.As well as androgens and estradiol.Androgens stimulate the secretion of estrogen and progesterone.The most important function of estradiol is to promote the formation of blood vessels in the second trimester,while progesterone plays an important role in early embryo implantation.Therefore,how to improve pregnancy success rate The main problems in the application of reproductive technology are closely related to the study of corpus luteum function.Steroid hormones dominate the formation and degeneration of the corpus luteum,but the exact molecular mechanism by which it interacts with lncRNA to regulate hormone secretion is still unclear.In recent years,the role of antisense long non-coding RNA in the field of reproduction has been reported,such as spermatogenesis,oocyte,and embryo development,follicular development and ovulation,and placenta formation.The results of previous studies in our laboratory showed that lncRNA Hand2os1/Uphwas significantly highly expressed in the corpus luteum on the 4 th and 18 th day of pregnancy.Hand2os1 is located on the antisense strand of Hand2（heart and neural crest derivatives-expressed transcript 2）.Hand2,as an important transcription factor downstream of the P₄ synthesis pathway,is in a pathway-dependent relationship with the PR receptor.Therefore,based on the above research,this article builds the early pregnancy corpus luteum cell model,after Hand2os1 is overexpressed or silenced,using RNAscope,CCK8,ELISA,and RT-q PCR technology to explore the lncRNA Hand2os1 in the mouse early pregnancy corpus luteum The expression,regulation and biological functions of corpus luteum;use in vitro hormone treatment to construct luteal cell models during estrus and degeneration,clarify the regulatory effect of Hand2os1 in the corpus luteum formation and degeneration stage,and lay the foundation for revealing the molecular mechanism of Hand2os1 regulating mouse corpus luteum formation and degeneration..The main research results obtained are as follows:（1）In early pregnancy mouse luteal cells,Hand2os1 is mainly located in the cytoplasm;overexpression or interference with Hand2os1 expression has no significant effect on the proliferation of luteal cells;（2）Overexpression or interference of Hand2os1 has no significant effect on the expression of Hand2.Overexpression and silencing of Hand2os1 will affect the secretion level of P₄.In-depth studies have found that it has a regulatory effect on the key genes Cyp11a1,3β-HSD,and St AR for steroid secretion,and is dependent on the St AR pathway;（3）Periodic corpus luteum model and in vitro experimental results show that Hand2os1expression has a downward trend during luteinization of granulosa cells in sexually mature mice,Hand2 m RNA expression is not affected,and Hand2os1 overexpression does not affect the secretion of P₄ during luteinization.And it has no significant effect on the expression of Cyp19a1 and St AR m RNA,the key factors of the luteinization process;（4）Overexpression of Hand2os1 has a certain promoting effect on PGF2α-induced corpus luteum degeneration.The apoptosis of luteal cells is also promoted but not significantly.The p-IRE1 and p-EIF2S1 signaling pathways related to endoplasmic reticulum stress are positively activated.The phenomenon,suggesting that Hand2os1 may participate in corpus luteal cell apoptosis by mediating endoplasmic reticulum stress;（5）Transcriptome GO function analysis and KEGG pathway enrichment results show that the signal pathways related to Hand2os1 regulating the function of luteal cells during pregnancy mainly include AMPK signal pathway,calcium signal pathway,and Hippo signal pathway;the biological functions involved include Protein transport in the plasma membrane,regulation of cytoplasmic calcium ion concentration,regulation of signal transduction receptor activity,etc.;（6）Hand2os1 may regulate ovarian function in early pregnancy mice by regulating the expression of related genes in the AMPK signaling pathway,Hippo signaling pathway,and calcium signaling pathway.In summary,Hand2os1 is specifically located in the cytoplasm of mouse luteal cells in early pregnancy.Hand2os1 plays a role in the P₄ secretion process by regulating the expression of steroid secretion marker genes St AR and Cyp11a1;Hand2os1 can cause P₄ levels by regulating the expression of St AR.Besides,Hand2os1 plays an insignificant biological role in the corpus luteum formation stage,but it stimulates the p-IRE1 and p-EIF2S1 pathways in endoplasmic reticulum stress through positive feedback during the corpus luteum degeneration stage and promotes cell apoptosis,which indicates that Hand2os1 plays an important role in the maintenance and degeneration of mouse corpus luteum function.In summary,Hand2os1is specifically located in the cytoplasm of mouse luteal cells in early pregnancy.Hand2os1plays a role in the P₄ secretion process by regulating the expression of steroid secretion marker genes St AR and Cyp11a1;Hand2os1 can cause P₄ levels by regulating the expression of St AR.Also,Hand2os1 has an insignificant biological role in the corpus luteum formation stage,but it stimulates the p-IRE1 and p-EIF2S1 pathways in endoplasmic reticulum stress through positive feedback during the corpus luteum degeneration stage to promote cell apoptosis,indicating that Hand2os1 plays an important role in the maintenance and degeneration of the mouse corpus luteum.