S1P Increases VEGF Production in Osteoblasts and Facilitates Endothelial Progenitor Cell Angiogenesis by Inhibiting miR-16-5p Expression via the c-Src/FAK Signaling Pathway in Rheumatoid Arthritis

Abstract

Angiogenesis is a critical process in the formation of new capillaries and a key participant in rheumatoid arthritis (RA) pathogenesis. Vascular endothelial growth factor (VEGF) stimulation of endothelial progenitor cells (EPCs) facilitates angiogenesis and the progression of RA. Phosphorylation of sphingosine kinase 1 (SphK1) produces sphingosine-1-phosphate (S1P), which increases inflammatory cytokine production, although the role of S1P in RA angiogenesis is unclear. In this study, we evaluated the impact of S1P treatment on VEGF-dependent angiogenesis in osteoblast-like cells (MG-63 cells) and the significance of SphK1 short hairpin RNA (shRNA) on S1P production in an in vivo model. We found significantly higher levels of S1P and VEGF expression in synovial fluid from RA patients compared with those with osteoarthritis by ELISA analysis. Treating MG-63 cells with S1P increased VEGF production, while focal adhesion kinase (FAK) and Src siRNAs and inhibitors decreased VEGF production in S1P-treated MG-63 cells. Conditioned medium from S1P-treated osteoblasts significantly increased EPC tube formation and migration by inhibiting miR-16-5p synthesis via proto-oncogene tyrosine-protein kinase src (c-Src) and FAK signaling in chick chorioallantoic membrane (CAM) and Matrigel plug assays. Infection with SphK1 shRNA reduced angiogenesis, articular swelling and cartilage erosion in the ankle joints of mice with collagen-induced arthritis (CIA). S1P appears to have therapeutic potential in RA treatment.

VEGF-A and FGF4 Engineered C2C12 Myoblasts and Angiogenesis in the Chick Chorioallantoic Membrane

“…A strong and potent angiogenic effect is seen in this study as a result of conditioned media taken from the stable transfected Myo-VEGF-GFP and Myo-FGF4-VEGF cell lines. This is consistent with other studies where conditioned media from osteoblast-like cells which secreted elevated levels of VEGF-A also induced a positive angiogenic response in the CAM assay [ 86 ].…”

Bioinformatics and System Biology Approach to Identify the Influences of COVID-19 on Rheumatoid Arthritis

“…Mir-26b-5p might be a novel biomarker for stratification of RA patients and was worth further clinical validation ( 76 ). Expression of mir-16-5p was inhibited, vascular endothelial growth factor (VEGF) production in osteoblasts is increased, and angiogenesis of endothelial progenitor cells is inhibited in RA ( 77 ). The results of Tao et al.…”

Microvascular Experimentation in the Chick Chorioallantoic Membrane as a Model for Screening Angiogenic Agents including from Gene-Modified Cells

“…Methods of adjusting the angiogenic potential of cells can vary from pre-treating cells with drugs or inflammatory factors such as sphingosine-1-phosphate [ 193 ], interferon or tumour necrosis factor [ 196 ], to more complex methods of genetically modifying the angiogenic behaviour of a cell.…”