Hypoxia Upregulates NOTCH3 Signaling Pathway to Promote Endothelial-Mesenchymal Transition in Pulmonary Artery Endothelial Cells

Abstract

Background. To investigate the effect of hypoxia on pulmonary artery endothelial cells and the role of NOTCH3 in endothelial-mesenchymal transition (EnMT) and to provide a research model for pulmonary disease and explain the pathogenesis of the pulmonary disease. Methods. Pulmonary artery endothelial cells were divided into two groups and cultured in normoxic and hypoxic environments, respectively. QPCR, western blot, and immunofluorescence were used to detect endothelial cell-specific marker protein and mRNA expression in each group, and the ability of endothelial cells migration was evaluated by scratch and transwell experiment. Results. The pulmonary artery endothelial cells in the normoxic group presented a typical pebble-like arrangement, and the endothelial cells in hypoxic culture showed a long spindle appearance. Hypoxia induced high expression of NOTCH3, Jagged-1, Hes1, c-Src, and CSL. Immunofluorescence showed that endothelial cells in hypoxic culture began to express the α-SMA, and the expression of vWF increased with hypoxia. Cell viability, scratch, and transwell results showed that endothelial cells in the hypoxic group were more capable of viability and migration than those in the normoxic group. The induction of EnMT by hypoxia can be inhibited by using notch3-specific inhibitor DAPT and Jagged-1. This study also found that miR-7-5p can regulate endothelial NOTCH3, indicating that miRNA is also involved in the process of endothelial-mesenchymal transformation. Conclusion. Hypoxia promotes the transformation of endothelial cells into mesenchymal cells by opening the NOTCH3 pathway, which lays the foundation for disease progression or clinical prognosis, and is of great significance in the treatment of diseases.

Notch3/Hes5 Induces Vascular Dysfunction in Hypoxia-Induced Pulmonary Hypertension Through ER Stress and Redox-Sensitive Pathways

“…This may involve upregulation of α-SMA (α-smooth muscle actin), a Notch3 signaling procontractile target. 30,31 Hypoxia is a potent inducer of oxidative stress 32 as observed in our study. Hypoxic alterations to vascular reactivity improved with fasudil and 4-PBA, suggesting a role for ROCK and ER stress in PH vascular dysfunction.…”

“…This may involve upregulation of α-SMA (α-smooth muscle actin), a Notch3 signaling procontractile target. 30 , 31 …”

Neural stem cell–derived exosomes regulate cell proliferation, migration, and cell death of brain microvascular endothelial cells via the miR‐9/Hes1 axis under hypoxia

“…Previous studies have revealed that Hes5 and Hes1 regulated the remodeling of blood vessels and the arterial specification of endothelial cells in brain vascular development 28 . Hypoxia caused a high expression of Notch3 and Hes1 and upregulated the Notch3 pathway to promote endothelial–mesenchymal transition in pulmonary artery endothelial cells (PAECs) 29 . The inhibition of Hes1 might play a protective role in endothelial cells under cholesterol stimulation via the PI3K/AKT signaling pathway 30 .…”

“… 28 Hypoxia caused a high expression of Notch3 and Hes1 and upregulated the Notch3 pathway to promote endothelial–mesenchymal transition in pulmonary artery endothelial cells (PAECs). 29 The inhibition of Hes1 might play a protective role in endothelial cells under cholesterol stimulation via the PI3K/AKT signaling pathway. 30 The Notch1 expression and transcriptional activity increased under hypoxic conditions in human PAECs (hPAECs).…”

Kidney Involvement in Systemic Sclerosis

“…In this permissive genetic background, abnormalities of the innate and adaptive immune systems, coupled with increased expression of ET-1 [26] and reduced production of nitric oxide synthase [27], cause vasoconstriction, intimal proliferation and hypoxia with consequent endothelial-mesenchymal transition and conversion to fibroblast-like cells [28]. In parallel, ET-1 activates and re-programs the functional phenotypes of vascular smooth muscle cells, microvascular pericytes and tissue fibroblasts into pro-fibrogenic cell In a predisposed genetic background, an exogenous trigger that targets blood vessels may determine the engagement of inflammatory molecules and cells of the innate immune system.…”