title: Hydrogen Peroxide and Lipopolysaccharide Differentially Affect the Expression of MicroRNAs 10a, 33a, 21, 221 in Endothelial Cells Before and After Coculture With Monocytes.
abstract: Inflammation and oxidative stress are important risk factors affecting various cells in the formation of atherosclerosis. MicroRNAs (miRs) are regulators of inflammation and atherogenesis. The expressions of endothelial cell (EC)-specific miR-10a and miR-21 and monocyte-specific miR-33a and miR-221 were investigated using coculture of the ECs and monocytes upon exposure to H2O2as an oxidative stressor, and endotoxin/lipopolysaccharide (LPS) as a microbial stressor. Human umbilical endothelial cells (HUVECs) and peripheral blood mononuclear cells (or monocytes) were cocultured in M199 complete medium and were incubated with LPS (20 ng/mL) or H2O2(1%) for 8 hours at 37°C. The HUVECs and monocytes were then separated from the cellular mix using a magnetic bead negative selection technique. The relative expression of miRs was determined by real-time polymerase chain reaction. In both cell types, H2O2induced miR10a ( P = 0.05) and LPS induced miR21 ( P = 0.0003) compared to the untreated controls. Coculture increased miR-10a and miR-21 expression in monocytes ( P = 0.0008 and <0.0001); however when cultured alone, HUVECs expressed higher levels of miR-10a and miR-21 ( P < 0.0001 and <0.0001). Coculture decreased the expression of miR-33a in monocytes ( P < 0.0001) while increasing miR221 in HUVECs and monocytes ( P < 0.0001 and <0.0001). The expression pattern of miRs in HUVECs and monocytes changes in the coculture compared to culturing alone in response to oxidative and microbial toxic compounds. Moreover, different cellular stressors induce different athero-miRs, which may affect the course of inflammation.
authors.affiliation: 1 Memory T Cell Laboratory, Department of Immunology, School of Medicine, Shiraz University of Medical Science, Shiraz, Iran.
authors.forename: Atefe Ghamar
authors.initials: AG
authors.lastname: Talepoor
authors.affiliation: 1 Memory T Cell Laboratory, Department of Immunology, School of Medicine, Shiraz University of Medical Science, Shiraz, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Kalani
authors.affiliation: 2 Department of Obstetrics and Gynecology, School of Medicine, Shiraz University of Medical Science, Shiraz, Iran.
authors.forename: Alamtaj Samsami
authors.initials: AS
authors.lastname: Dahaghani
authors.affiliation: 1 Memory T Cell Laboratory, Department of Immunology, School of Medicine, Shiraz University of Medical Science, Shiraz, Iran.
authors.forename: Mehrnoosh
authors.initials: M
authors.lastname: Doroudchi
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D008070:Lipopolysaccharides / Q000633:toxicity*; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism
chemical_list: D008070:Lipopolysaccharides; D035683:MicroRNAs; D006861:Hydrogen Peroxide
keywords: H2O2; HUVEC; coculture; endotoxin/LPS; microRNA; monocyte