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Fatemeh Roodbari

Fatemeh Roodbari

Academic rank: Assistant Professor
ORCID:
Education: PhD.
ScopusId:
Faculty: Science
Address: University of Mazandaran
Phone: 01135302424

Research

Title
Activation of toll‑like receptor signaling in endothelial progenitor cells dictates angiogenic potential: from hypothesis to actual stat
Type
JournalPaper
Keywords
Human endothelial progenitor cells · Toll-like receptor signaling pathway · Lipopolysaccharide · Maturation · Angiogenic capacit
Year
2021
Journal CELL AND TISSUE RESEARCH
DOI
Researchers Morteza Heidarzadeh ، Çığır Biray Avcı ، Shirin Saberianpour ، Mahdi Ahmadi ، Mehdi Hassanpour ، Rezai Rahbarghazi ، Hesam Saghaei Bagheri ، Mehdi Talebi ، Jafar Rezaie ، Masoud Darabi ، Emel Sokulu ، Fatemeh Roodbari

Abstract

Human endothelial progenitor cells (EPCs) were isolated from cord blood samples and enriched by magnetic activated cell sorting method based on the CD133 marker. Cells were incubated with diferent doses of bacterial lipopolysaccharide, ranging from 2, 5, 10, 50, 100, 200, 250, 500, to 1000 µg/ml, for 48 h. The cell survival rate was determined by using MTT assay. To confrm activation of the toll-like receptor signaling pathway, PCR array analysis was performed. Protein levels of ERK1/2, p-ERK1/2, NF-ƙB and TRIF proteins were measured using western blotting. The content of TNF-α and lipoprotein lipase activity were analyzed by immunofuorescence imaging. Flow cytometric analysis of CD31 was performed to assess the maturation rate. Cell migration was studied by the Transwell migration assay. The expression of genes related to exosome biogenesis was measured using real-time PCR analysis. In vivo gel plug angiogenesis assay was done in nude mice. Lipopolysaccharide changed endothelial progenitor cells’ survival in a dose-dependent manner with maximum viable cells in groups treated with 2 µg/ml. PCR array analysis showed the activation of toll-like signaling pathways after exposure to LPS (p<0.05). Western blotting analysis indicated an induction of p-ERK1/2 and Erk1/2, NF-kB and TRIF in LPS-treated EPCs compared with the control (p<0.05). Immunofuorescence staining showed an elevation of TNF-α and lipoprotein lipase activity after lipopolysaccharide treatment (p<0.05). Lipopolysaccharide increased EPC migration and expression of exosome biogenesis-related genes (p<0.05). In vivo gel plug analysis revealed enhanced angiogenesis in cells exposed to bacterial lipopolysaccharide. Data highlighted the close relationship between the toll-like receptor signaling pathway and functional activity in EPCs.