caa a22 4500 475773985 CHVBK 20180406123625.0 cr unu---uuuuu 170329e20000701xx s 000 0 eng 10.1007/s004180000160 doi (NATIONALLICENCE)springer-10.1007/s004180000160 NADPH oxidase subunits and superoxide production in porcine pulmonary artery endothelial cells [Elektronische Daten] [B. Höhler, B. Holzapfel, W. Kummer] Abstract.: An NADPH oxidase complex composed of a membrane-bound flavocytochromeb558 consisting of two subunits (p22phox and gp91phox) and cytosolic activating factors (p47phox and p67phox) generates superoxide anions from oxygen in the respiratory burst of phagocytic cells. Inconsistent results have been previously obtained concerning its additional occurrence in pulmonary artery endothelial cells (PAEC), and this issue was addressed in the present study. PAEC isolated from porcine pulmonary trunk contained mRNA for p22phox and gp91phox as demonstrated by reverse transcription-polymerase chain reaction. Immunohistochemistry demonstrated cytochrome subunits, p22phox, gp91phox, p47phox, and p67phox, both in vitro in isolated PAEC and in situ in endothelial cells in tissue sections of the pulmonary trunk. Isolated PAEC generated reactive oxygen species (ROS; measured by lucigenin-induced chemiluminescence and conversion of dihydrorhodamine123 into rhodamine123) in response to stimulation with phorbol 12-myristate 13-acetate. This stimulated ROS production was sensitive to the flavoprotein inhibitor diphenyleneiodonium, and reduced when the superoxide scavenger superoxide dismutase was added. Chemiluminescence measurements of superoxide generation by stimulated PAEC accounted for approximately 1% of that generated by stimulated peritoneal macrophages. The data demonstrate a low-output NADPH oxidase system in porcine PAEC sharing several components with that identified in phagocytic cells. Springer-Verlag, 2000 Cytochromeb558 Endothelium NADPH oxidase Pulmonary artery Smooth muscle cells Superoxide anion Pig nationallicence Höhler B. Institute for Anatomy and Cell Biology, Justus-Liebig-University, Aulweg 123, 35385 Giessen, Germany aut Holzapfel B. Institute for Anatomy and Cell Biology, Justus-Liebig-University, Aulweg 123, 35385 Giessen, Germany aut Kummer W. Institute for Anatomy and Cell Biology, Justus-Liebig-University, Aulweg 123, 35385 Giessen, Germany aut https://doi.org/10.1007/s004180000160 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s004180000160 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Höhler B. Institute for Anatomy and Cell Biology, Justus-Liebig-University, Aulweg 123, 35385 Giessen, Germany aut NATIONALLICENCE 700 1- Holzapfel B. Institute for Anatomy and Cell Biology, Justus-Liebig-University, Aulweg 123, 35385 Giessen, Germany aut NATIONALLICENCE 700 1- Kummer W. Institute for Anatomy and Cell Biology, Justus-Liebig-University, Aulweg 123, 35385 Giessen, Germany aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer