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Melatonin Regulates the Expression of VEGF and HOXA10 in Bovine Endometrial Epithelial Cells through the SIRT1/PI3K/AKT Pathway

文献类型： 外文期刊

作者： Li, Qi;Tang, Ying;Chen, Yanru;Li, Bo;Wang, Hongzhan;Liu, Shicheng;Adeniran, Samson O.;Zheng, Peng

作者机构：

关键词： endometrial epithelial cells;melatonin;PI3K;VEGF;HOXA10

期刊名称： ANIMALS

ISSN： 2076-2615

年卷期： 2024 年 14 卷 19 期

页码：

收录情况： SCIE（2024版）

摘要： Simple Summary The low embryo attachment rate and early embryo loss of dairy cows are important factors restricting the improvement of dairy cow fertility and are also the key scientific problems that need to be solved urgently in dairy cow production and embryo transfer. In this study, we used RT-qPCR, Western blot, immunofluorescence, and so on in cultured bovine endometrial epithelial cells to investigate the mechanism by which melatonin regulates the expression of VEGF and HOXA10 through SIRT1, PI3K, AKT, and miRNA. The results indicate that melatonin regulates the expression of VEGF and HOXA10 through the SIRT1/PI3K/AKT pathway and promotes the establishment of receptivity in bovine endometrial epithelial cells. The results indicate that melatonin negatively regulates the expression of bta-miR-497 and bta-27a-3p in bovine endometrial epithelial cells through the SIRT1/PI3K/AKT pathway and positively regulates the expression of VEGF and HOXA10. In this regulatory network, bta-miR-27a-3p negatively modulates HOXA10 expression, while bta-miR-497 negatively modulates VEGF expression.Abstract Melatonin plays a critical role in regulating embryo attachment in ruminants. While numerous studies have investigated its effects on early embryo development in vitro, the precise mechanisms by which melatonin influences the receptivity of endometrial epithelial cells in dairy cows remain unclear. The prerequisite for embryo implantation is the specific physiological condition of the endometrium that allows the embryo to implant, also known as endometrial receptivity. In addition to this, endometrial cells undergo processes such as proliferation, differentiation, and renewal, which makes the embryo more easily implanted. In this study, bovine endometrial epithelial cells were cultured and treated with melatonin, Silent Information Regulator 1 (SIRT1) inhibitor (EX527), and protein kinase B (AKT) phosphorylation inhibitor (periposine). RT-qPCR, Western blot, and immunofluorescence analysis were performed to investigate the effects of melatonin on the expression of target gene (SIRT1); cell proliferative genes, phosphatidylinositol-4,5-bisphosphate 3-Kinase (PI3K), AKT, cyclinD1, cyclinE1; and receptive genes (Leukemia Inhibitory Factor (LIF), Vascular Endothelial Growth Factor (VEGF), Homeobox Structure Gene 10 (HOXA10)). Additionally, microRNA (miRNA) mimics and inhibitors were used to transfect the cells to study the regulatory relationship between miRNA and receptive genes. Results indicated that melatonin activates the PI3K/AKT signaling pathway, upregulates cyclinD1 and cyclinE1, and promotes the proliferation of bovine endometrial epithelial cells. Melatonin also upregulated the expression of VEGF and HOXA10 and downregulated the expression of bta-miR-497 and bta-miR-27a-3p through SIRT1/PI3K/AKT signaling pathway. Further, bta-miR-497 and bta-miR-27a-3p were found to negatively regulate VEGF and HOXA10, respectively. Therefore, melatonin regulates the expression of VEGF and HOXA10 through the SIRT1/PI3K/AKT pathway and promotes the establishment of receptivity in bovine endometrial epithelial cells.

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