title: Double-stranded RNA attenuates the barrier function of human pulmonary artery endothelial cells.
abstract: Circulating RNA may result from excessive cell damage or acute viral infection and can interact with vascular endothelial cells. Despite the obvious clinical implications associated with the presence of circulating RNA, its pathological effects on endothelial cells and the governing molecular mechanisms are still not fully elucidated. We analyzed the effects of double stranded RNA on primary human pulmonary artery endothelial cells (hPAECs). The effect of natural and synthetic double-stranded RNA (dsRNA) on hPAECs was investigated using trans-endothelial electric resistance, molecule trafficking, calcium (Ca(2+)) homeostasis, gene expression and proliferation studies. Furthermore, the morphology and mechanical changes of the cells caused by synthetic dsRNA was followed by in-situ atomic force microscopy, by vascular-endothelial cadherin and F-actin staining. Our results indicated that exposure of hPAECs to synthetic dsRNA led to functional deficits. This was reflected by morphological and mechanical changes and an increase in the permeability of the endothelial monolayer. hPAECs treated with synthetic dsRNA accumulated in the G1 phase of the cell cycle. Additionally, the proliferation rate of the cells in the presence of synthetic dsRNA was significantly decreased. Furthermore, we found that natural and synthetic dsRNA modulated Ca(2+) signaling in hPAECs by inhibiting the sarco-endoplasmic Ca(2+)-ATPase (SERCA) which is involved in the regulation of the intracellular Ca(2+) homeostasis and thus cell growth. Even upon synthetic dsRNA stimulation silencing of SERCA3 preserved the endothelial monolayer integrity. Our data identify novel mechanisms by which dsRNA can disrupt endothelial barrier function and these may be relevant in inflammatory processes.
authors.affiliation: Department of Anesthesia and Intensive Care Medicine, Medical University of Graz, Graz, Austria.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Bálint
authors.forename: Diana
authors.initials: D
authors.lastname: Zabini
authors.forename: Viktoria
authors.initials: V
authors.lastname: Konya
authors.forename: Chandran
authors.initials: C
authors.lastname: Nagaraj
authors.forename: Attila G
authors.initials: AG
authors.lastname: Végh
authors.forename: György
authors.initials: G
authors.lastname: Váró
authors.forename: Imola
authors.initials: I
authors.lastname: Wilhelm
authors.forename: Csilla
authors.initials: C
authors.lastname: Fazakas
authors.forename: István A
authors.initials: IA
authors.lastname: Krizbai
authors.forename: Akos
authors.initials: A
authors.lastname: Heinemann
authors.forename: Horst
authors.initials: H
authors.lastname: Olschewski
authors.forename: Andrea
authors.initials: A
authors.lastname: Olschewski
mesh_terms: D000199:Actins / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003911:Dextrans / Q000378:metabolism; D017097:Electric Impedance; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D006632:Histamine / Q000494:pharmacology; D006706:Homeostasis / Q000187:drug effects; D006801:Humans; D007365:Intercellular Junctions / Q000187:drug effects / Q000378:metabolism; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011070:Poly I-C / Q000494:pharmacology; D011651:Pulmonary Artery / Q000166:cytology*; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000378:metabolism; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism; D000099211:Phospholamban
chemical_list: D000199:Actins; D002118:Calcium; D002135:Calcium-Binding Proteins; D003911:Dextrans; D016650:Fluorescein-5-isothiocyanate; D006632:Histamine; D019869:Phosphatidylinositol 3-Kinases; D011070:Poly I-C; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000099211:Phospholamban; C015219:fluorescein isothiocyanate dextran