| Objective:The primary purpose of this study is to investigate the role of LAMA4 in trophoblast and human umbilical vein endothelial cells (HUVECs) function during the development of PE.Methods:Here we show that LAMA4 was expressed in the first trimester and term human placentas by immunohistochemistry. LAMA4 siRNA significantly inhibited outgrowth of extravillous explants in vitro, as well as invasion and migration of HTR-8/SVneo cells; migration and tube formation of HUVECs. HTR-8/SVneo cells, HUVECs and extravillous explants were used to mimic the effects of a Hypoxia-Reoxygenation (H/R) environment on placentas to study LAMA4 expression and trophoblast or endothelial cells fuctions. Inhibitor of p38 MAPK signaling pathway was used to study the internal mechanism of LAMA4 in HTR-8/SVneo and HUVEC cells.Results:Our study shows that LAMA4 protein levels in pre-eclampsia were significantly lower as compared to the control placentas. LAMA4 was highly expressed in trophoblast and human endothelial cells during the first trimester. H/R condition and LAMA4 siRNA decreased the invasive and migratory abilities of cells-decreased the expression of MMP2 and MMP9 in consistent with increased TEMPI and TIMP2. The LAMA4 small-interfering RNA transfection and hypoxia-reoxygenation (H/R) intervention reduced the migratory and tube formation abilities of HUVECs. The mitogen-activated protein kinase (MAPK) signaling pathways interacted with LAMA4 expression and H/R exposure led to MAPK pathways activation in trophoblast and HUVECs. Furthermore, silencing of LAMA4 resulted in an altered expression of proteins associated with the p38 signaling pathway.Conclsion:Our results suggest that LAMA4 promotes human trophoblast cell invasion, migration and endothelial cells angiogenesis. Oxidative stress plays a vital role in controlling expression of LAMA4 through MAPK signaling pathways, which suggests a possible pathological mechanism of PE. |
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