Abstract
Formation and limited lifespan of corpus luteum (CL) are important for proper ovarian periodicity and fertility. Failed vascularization, imbalance between proliferation and apoptosis leads to luteal phase deficiency and infertility. The aim of this study was to examine the effect of vaspin on angiogenesis, apoptosis and proliferation as well as the involvement of 78-kDa glucose-regulated protein receptor (GRP78) and mitogen-activated kinase (MAP3/1) in these processes. Porcine luteal cells were incubated with vaspin (0.1–10 ng/mL) for 24 h to 72 h and then mRNA and protein expression of angiogenesis: vascular endothelial growth factor (VEGFA), fibroblast growth factor 2 (FGF2), angiopoietin 1 (ANGPT1), VEGFA receptors (VEGFR1, VEGFR2), apoptosis: caspase 3, bcl-2-like protein 4 (BAX), B-cell lymphoma (BCL2), and proliferation: proliferating cells nuclear antigen (PCNA), cyclin A factors as well as secretion of VEGFA, FGF2, ANGT1 were measured by real-time polymerase chain reaction (PCR), immunoblotting and enzyme-linked immunosorbent assay (ELISA), respectively. Moreover, apoptosis was assessed by caspase activity using the Caspase-Glo 3/7 assay, while proliferation was by alamarBlue. We found that vaspin enhanced luteal cell angiogenesis, proliferation, and significantly decreased apoptosis. Additionally, using GRP78 siRNA and the pharmacological inhibitor of MAP3/1 (PD98059), we observed that the effect of vaspin was reversed to the control level in all investigated processes. Taken together, our results suggest that vaspin is a new regulator of female fertility by direct regulation of CL formation and maintenance of luteal cell function.
Highlights
Formation and limited lifespan of corpus luteum (CL) is important for proper ovarian periodicity and fertility [1]
Dose-Dependent Effect of Vaspin on Luteal Cell Angiogenesis Vaspin is linked with angiogenesis in human portal tracts and lobules in chronic hepatitis liver [22]
We hypothesize that the stimulatory effect of vaspin on luteal angiogenesis is through the direct action of vaspin on mRNA expression of angiogenic factors, independent of the properties of mitogenic vaspin
Summary
Formation and limited lifespan of corpus luteum (CL) is important for proper ovarian periodicity and fertility [1]. Inappropriate vascularization leads to aberrant CL development and reduces the serum concentration of P4 [3], causing miscarriages [4]. Angiogenesis is regulated by several factors including vascular endothelial growth factor (VEGFA), fibroblast growth factor 2 (FGF2), and angiopoietin 1 (ANGPT1). Both VEGFA and FGF2 promotes endothelial cell differentiation, proliferation, migration, and vascular tube formation [5], while ANGPT1 promotes the maturation and stabilization of nascent vessels [6]. The aims of the research are to better understand the processes of CL establishment, formation, angiogenesis, and regression as well as molecular mechanisms in luteal cells function. The obtained data will contribute to solving the problem of infertility caused by CL failure
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