Reactive oxygen species production via NADPH oxidase mediates TGF-beta-induced cytoskeletal alterations in endothelial cells.
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.
Hu
Taishan
T
The Dorrance Hamilton Research Laboratories, Thomas Jefferson Univ., Philadelphia, PA 19107, USA.
Ramachandrarao
Satish P
SP
Siva
Senthuran
S
Valancius
Cathryn
C
Zhu
Yanqing
Y
Mahadev
Kalyankar
K
Toh
Irene
I
Goldstein
Barry J
BJ
Woolkalis
Marilyn
M
Sharma
Kumar
K
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
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