Reactive oxygen species production via NADPH oxidase mediates TGF-β-induced cytoskeletal alterations in endothelial cells

Hu, Tai‐Shan; RamachandraRao, Satish P.; Senthuran, Siva; Valancius, Cathryn; Zhu, Yanqing; Mahadev, Kalyankar; Toh, Irene; Goldstein, Barry J.; Woolkalís, Marílyn J.; Sharma, Kumar

doi:10.1152/ajprenal.00024.2005

Cited by 157 publications

(116 citation statements)

References 44 publications

(46 reference statements)

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“…33,49 Because Nox4 was increased in Dcn Ϫ/Ϫ kidneys even in the absence of diabetes, the data suggest that decorin may have a direct effect in regulating Nox4 expression. The effect on Nox4 may well be related to TGF-␤, given our recent demonstrations that TGF-␤-induced reactive oxygen species production in endothelial cells is mediated by Nox-4 50 and that TGF-␤ can stimulate Nox4 production. 51 Nox4 was found to be expressed in both mesangial cells and podocytes, suggesting that increased oxidant tone in these cell types contribute to cell dysfunction.…”

Section: Discussionmentioning

confidence: 96%

Decorin Deficiency Enhances Progressive Nephropathy in Diabetic Mice

Williams

Qiu

Usui

et al. 2007

The American Journal of Pathology

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Decorin, a proteoglycan that inhibits active transforming growth factor-␤, is increased in diabetic nephropathy; however, its functional significance is unclear. In this study, we used low-dose streptozotocin to induce type 1 diabetes in wild-type (C57BL/6J Dcn ؉/؉ ), Dcn ؊/؊ , and Dcn ؉/؊ mice and studied the mice for up to 1 year of diabetes. Decorin gene dose had no effect on severity of diabetes; however, the Dcn ؊/؊ diabetic mice died significantly earlier than nondiabetic controls (57 versus 7.3% mortality). In contrast to wild-type diabetic mice, which failed to develop significant nephropathy, the Dcn ؊/؊ diabetic mice developed a significant increase in albuminuria and plasma creatinine and a concurrent decrease in circulating adiponectin levels. Interestingly, adiponectin levels at 6 months of diabetes were predictive of mortality in diabetic mice.

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Section: Discussionmentioning

confidence: 96%

Decorin Deficiency Enhances Progressive Nephropathy in Diabetic Mice

Williams

Qiu

Usui

et al. 2007

The American Journal of Pathology

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“…Accumulating evidence supports a link between the actin cytoskeleton and actin-binding proteins in the activation of phagocytic and non-phagocytic NADPH oxidase (23)(24)(25)(26)(27). Stimulation of neutrophils with phorbol ester induces translocation of p47 phox to the cytoskeleton without altering the distribution of either p40 phox or p67 phox and increases the oxidase activity that co-sediments with a heavy plasma membrane fraction consisting of actin and fodrin (28,29).…”

Section: Discussionmentioning

confidence: 97%

“…Actin cytoskeleton and other cytoskeletal proteins may play a role in phagocytic and non-phagocytic assembly and the activation of NADPH oxidase (23)(24)(25)(26)(27). In phorbol ester-stimulated neutrophils, oxidase activity has been shown to co-sediment with the heavy plasma membrane fraction that contains actin and fodrin; furthermore, the labile oxidase can be stabilized by chemical cross-linking and is not extractable by Triton X-100, suggesting that an interaction occurs between the NADPH oxidase complex and actin filaments (28,29).…”

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confidence: 99%

Regulation of Hyperoxia-induced NADPH Oxidase Activation in Human Lung Endothelial Cells by the Actin Cytoskeleton and Cortactin

Usatyuk

Romer

et al. 2007

Journal of Biological Chemistry

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Although the actin cytoskeleton has been implicated in the control of NADPH oxidase in phagocytosis, very little is known about the cytoskeletal regulation of endothelial NADPH oxidase assembly and activation. Here, we report a role for cortactin and the tyrosine phosphorylation of cortactin in hyperoxiainduced NADPH oxidase activation and ROS production in human pulmonary artery ECs (HPAECs). Exposure of HPAECs to hyperoxia for 3 h induced NADPH oxidase activation, as demonstrated by enhanced superoxide production. Hyperoxia also caused a thickening of the subcortical dense peripheral F-actin band and increased the localization of cortactin in the cortical regions and lamellipodia at cell-cell borders that protruded under neighboring cells. Pretreatment of HPAECs with the actin-stabilizing agent phallacidin attenuated hyperoxia-induced cortical actin thickening and ROS production, whereas cytochalasin D and latrunculin A enhanced basal and hyperoxia-induced ROS formation. In HPAECs, a 3-h hyperoxic exposure enhanced the tyrosine phosphorylation of cortactin and interaction between cortactin and p47 phox , a subcomponent of the EC NADPH oxidase, when compared with normoxic cells. Furthermore, transfection of HPAECs with cortactin small interfering RNA or myristoylated cortactin Src homology domain 3 blocking peptide attenuated ROS production and the hyperoxia-induced translocation of p47 phox to the cell periphery. Similarly, down-regulation of Src with Src small interfering RNA attenuated the hyperoxia-mediated phosphorylation of cortactin tyrosines and blocked the association of cortactin with actin and p47phox . In addition, the hyperoxia-induced generation of ROS was significantly lower in ECs expressing a tyrosine-deficient mutant of cortactin than in vector control or wild-type cells. These data demonstrate a novel function for cortactin and actin in hyperoxiainduced activation of NADPH oxidase and ROS generation in human lung endothelial cells. production that is dependent on NADPH oxidase activation and independent of the mitochondrial electron transport or xanthine/xanthine oxidase systems (10). The mechanisms of NADPH oxidase activation are complex. In phagocytes, activation of NADPH oxidase requires serine phosphorylation of the cytosolic p47 phox , p67 phox , and p40 phox components, assembly of the phosphorylated subunits with Rac2, and translocation to the phagosomes for association with cytochrome b 558. Here, one-electron reduction of molecular O 2 to O 2 . occurs with NADPH as the electron donor (7). In leukocytes, formyl-Met-Leu-Phe-OH or phorbol ester stimulates phosphorylation of p47 phox at multiple serine residues through reactions involving several protein kinases such as protein kinase C, protein kinase A, and mitogen-activated protein kinases (14 -17). In HPAECs, tumor necrosis factor-␣-medi-*This work was supported by National Institutes of Health Grants RO1 HL 69909 (to V. N.), PO1 HL 58064 (to V. N. and J. G. N. G.) and DE13079-01 and AI061042 (to L. H. R.). The costs of publication...

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“…22 Because PFD has been postulated to inhibit NADPH oxidase in other cell types, 23 we determined whether there was a similar effect of PFD to block TGF-␤-induced ROS in mesangial cells. As shown in Figure 3, PFD blocked the TGF-␤-induced increase in the ROS production in mesangial cells (100 to 1000 g/ml, in a dosage-dependent manner).…”

Section: Pirfenidone Reduces Tgf-␤-induced Mesangial Cell Ros Generationmentioning

confidence: 99%

Pirfenidone Is Renoprotective in Diabetic Kidney Disease

RamachandraRao

Zhu

Ravasi

et al. 2009

Journal of the American Society of Nephrology

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148

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Although several interventions slow the progression of diabetic nephropathy, current therapies do not halt progression completely. Recent preclinical studies suggested that pirfenidone (PFD) prevents fibrosis in various diseases, but the mechanisms underlying its antifibrotic action are incompletely understood. Here, we evaluated the role of PFD in regulation of the extracellular matrix. In mouse mesangial cells, PFD decreased TGF-␤ promoter activity, reduced TGF-␤ protein secretion, and inhibited TGF-␤-induced Smad2-phosphorylation, 3TP-lux promoter activity, and generation of reactive oxygen species. To explore the therapeutic potential of PFD, we administered PFD to 17-wk-old db/db mice for 4 wk. PFD treatment significantly reduced mesangial matrix expansion and expression of renal matrix genes but did not affect albuminuria. Using liquid chromatography with subsequent electrospray ionization tandem mass spectrometry, we identified 21 proteins unique to PFD-treated diabetic kidneys. Analysis of gene ontology and protein-protein interactions of these proteins suggested that PFD may regulate RNA processing. Immunoblotting demonstrated that PFD promotes dosage-dependent dephosphorylation of eukaryotic initiation factor, potentially inhibiting translation of mRNA. In conclusion, PFD is renoprotective in diabetic kidney disease and may exert its antifibrotic effects, in part, via inhibiting RNA processing.

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Reactive oxygen species production via NADPH oxidase mediates TGF-β-induced cytoskeletal alterations in endothelial cells

Cited by 157 publications

References 44 publications

Decorin Deficiency Enhances Progressive Nephropathy in Diabetic Mice

Decorin Deficiency Enhances Progressive Nephropathy in Diabetic Mice

Regulation of Hyperoxia-induced NADPH Oxidase Activation in Human Lung Endothelial Cells by the Actin Cytoskeleton and Cortactin

Pirfenidone Is Renoprotective in Diabetic Kidney Disease

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