Hydrogen peroxide-mediated transcriptional induction of macrophage colony-stimulating factor by TGF-beta1.
TGF-beta1 and macrophages are important regulators of tissue fibrosis and remodeling. Here we show that TGF-beta1 induces the expression of macrophage-CSF (M-CSF) in vascular endothelial cells via a signaling pathway(s) involving hydrogen peroxide (H2O2). In a time-dependent manner, TGF-beta1 produced a 10- and a 6-fold increase in M-CSF mRNA and protein levels after 12 h, respectively. This increase in M-CSF expression was attenuated by a nitric oxide donor, S-nitrosoglutathione (GSNO), and by a nonspecific oxidase inhibitor, diphenylene iodonium. Furthermore, the TGF-beta1-induced M-CSF mRNA expression was inhibited by catalase, but not by superoxide dismutase, suggesting that H2O2 rather than superoxide anion (O2.-) is the primary mediator of the effects of TGF-beta1. Transient transfection studies using deletional M-CSF promoter constructs demonstrated that TGF-beta1 produced a 13-fold induction in M-CSF promoter activity that was repressed by >85% with GSNO and catalase, in part through inhibitory effects on kappaB cis-acting elements. Electrophoretic mobility shift assays revealed that the activation of nuclear factor-kappaB by TGF-beta1 was also inhibited by GSNO and catalase, but not by superoxide dismutase. In a concentration-dependent manner, treatment with exogenous H2O2 produced 14- and 4.6-fold increases in M-CSF promoter activity and mRNA expression, respectively. These results indicate that the generation of H2O2 and activation of NF-kappaB by TGF-beta1 are required for the induction of M-CSF gene transcription.
Hong
Y H
YH
Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
Peng
H B
HB
La Fata
V
V
Liao
J K
JK
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001713:Biphenyl Compounds / Q000494:pharmacology; D002374:Catalase / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D005978:Glutathione / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism* / Q000494:pharmacology; D016173:Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000235:genetics; D016328:NF-kappa B / Q000378:metabolism; D009603:Nitroso Compounds / Q000494:pharmacology; D009861:Onium Compounds / Q000494:pharmacology; D010088:Oxidoreductases / Q000037:antagonists & inhibitors; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis; D026422:S-Nitrosoglutathione; D015398:Signal Transduction / Q000502:physiology*; D013482:Superoxide Dismutase / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects*; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D000975:Antioxidants; D001713:Biphenyl Compounds; D004791:Enzyme Inhibitors; D016328:NF-kappa B; D009603:Nitroso Compounds; D009861:Onium Compounds; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; C031291:diphenyliodonium; D026422:S-Nitrosoglutathione; D016173:Macrophage Colony-Stimulating Factor; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; D002374:Catalase; D013482:Superoxide Dismutase; D005978:Glutathione