Abstract
Vascular endothelial growth factor receptor-2 (VEGFR-2) regulates the mitogen-activated protein kinase (MAPK) signaling pathway and plays an important role in angiogenesis. Bu Shen Zhu Yun decoction (BSZYD) can improve endometrial receptivity and embryo implantation rates in patients undergoing in vitro fertilization. However, whether BSZYD improves endometrial receptivity via angiogenesis remains unclear. Here, we investigated the effects of BSZYD on the proliferation, migration, and angiogenesis of human endometrial microvascular endothelial cells (HEMECs) and found that BSZYD upregulated the expression of cyclin D1, matrix metalloproteinase 9 (MMP9), and proliferating cell nuclear antigen (PCNA) in HEMECs. Cell Counting Kit 8 assay, scratch-wound assay, and Tube Formation Assay results showed that BSZYD promoted the proliferation, migration, and angiogenesis of HEMECs. Western blot analysis results revealed the activation of the MAPK signaling pathway by BSZYD through the upregulation of VEGF and VEGFR-2 expression. Together, these findings highlight the novel mechanism underlying BSZYD-mediated improvement in endometrial receptivity through the MAPK signaling pathway.
Highlights
Implantation is a critical step in pregnancy, and good endometrial receptivity is an important factor for implantation
We found that the expression levels of Vascular endothelial growth factor receptor-2 (VEGFR-2), proliferating cell nuclear antigen (PCNA), cyclin D1, and matrix metalloproteinase 9 (MMP9) increased in a time-dependent manner and reached peak values at 12 h (P < 0.05). e highest levels were maintained until 24 h (Figures 1(a) and 1(b))
Cell Counting Kit-8 (CCK-8) assay results showed that Vascular endothelial growth factor (VEGF) induced cell proliferation (Figure 1(c)). e results of the scratch-wound assay showed that wound healing was significantly faster in VEGF-stimulated cells than in untreated cells (Figure 1(d)), indicating that VEGF promotes human endometrial microvascular endothelial cells (HEMECs) migration
Summary
Implantation is a critical step in pregnancy, and good endometrial receptivity is an important factor for implantation. Embryo implantation is a continuous dynamic process, during which the womb environment undergoes a series of changes. The endometrium apical surface undergoes several morphological, molecular, and biochemical changes to provide a favorable environment for embryo implantation [1, 2]. Considering the periodic proliferation of endometrial vascular system during embryo implantation and development [4], endometrial angiogenesis is regulated by several vasoactive substances and angiogenic factors. Vascular endothelial growth factor receptor-2 (VEGFR-2/KDR) is considered as the most important VEGF receptor in the process of angiogenesis and known to regulate endometrial angiogenesis [6]. Mitogen-activated protein kinase (MAPK) signaling pathway, located downstream of VEGFR-2, can regulate the proliferation and migration of vascular endothelial cells by a series of cascade reactions [7]
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