title: Reactive oxygen species production via NADPH oxidase mediates TGF-beta-induced cytoskeletal alterations in endothelial cells.
abstract: Cytoskeletal alterations in endothelial cells have been linked to nitric oxide generation and cell-cell interactions. Transforming growth factor (TGF)-beta has been described to affect cytoskeletal rearrangement in numerous cell types; however, the underlying pathway is unclear. In the present study, we found that human umbilical vein endothelial cells (HUVEC) have marked cytoskeletal alterations with short-term TGF-beta treatment resulting in filipodia formation and F-actin assembly. The cytoskeletal alterations were blocked by the novel TGF-beta type I receptor/ALK5 kinase inhibitor (SB-505124) but not by the p38 kinase inhibitor (SB-203580). TGF-beta also induced marked stimulation of reactive oxygen species (ROS) within 5 min of TGF-beta exposure. TGF-beta stimulation of ROS was mediated by the NAPDH oxidase homolog Nox4 as DPI, an inhibitor of NADPH oxidase, and dominant-negative Nox4 adenovirus blocked ROS production. Finally, inhibition of ROS with ROS scavengers or dominant-negative Nox4 blocked the TGF-beta effect on cytoskeleton changes in endothelial cells. In conclusion, our studies show for the first time that TGF-beta-induced ROS production in human endothelial cells is via Nox4 and that TGF-beta alteration of cytoskeleton in HUVEC is mediated via a Nox4-dependent pathway.
authors.affiliation: The Dorrance Hamilton Research Laboratories, Thomas Jefferson Univ., Philadelphia, PA 19107, USA.
authors.forename: Taishan
authors.initials: T
authors.lastname: Hu
authors.forename: Satish P
authors.initials: SP
authors.lastname: Ramachandrarao
authors.forename: Senthuran
authors.initials: S
authors.lastname: Siva
authors.forename: Cathryn
authors.initials: C
authors.lastname: Valancius
authors.forename: Yanqing
authors.initials: Y
authors.lastname: Zhu
authors.forename: Kalyankar
authors.initials: K
authors.lastname: Mahadev
authors.forename: Irene
authors.initials: I
authors.lastname: Toh
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Goldstein
authors.forename: Marilyn
authors.initials: M
authors.lastname: Woolkalis
authors.forename: Kumar
authors.initials: K
authors.lastname: Sharma
mesh_terms: D000199:Actins / Q000096:biosynthesis; D030201:Activin Receptors, Type I / Q000502:physiology; D000256:Adenoviridae / Q000235:genetics; D002460:Cell Line; D003599:Cytoskeleton / Q000502:physiology*; D042783:Endothelial Cells / Q000502:physiology* / Q000648:ultrastructure; D006579:Heterozygote; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000378:metabolism* / Q000502:physiology; D017346:Protein Serine-Threonine Kinases; D017382:Reactive Oxygen Species / Q000378:metabolism*; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000502:physiology*; D048051:p38 Mitogen-Activated Protein Kinases / Q000502:physiology
chemical_list: D000199:Actins; D017382:Reactive Oxygen Species; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C498910:NOX4 protein, human; D017346:Protein Serine-Threonine Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D030201:Activin Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625841:TGFBR1 protein, human