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Type: Journal article
Title: TGF-β1-induced MAPK activation promotes collagen synthesis, nodule formation, redox stress and cellular senescence in porcine aortic valve interstitial cells
Other Titles: TGF-(beta)1-induced MAPK activation promotes collagen synthesis, nodule formation, redox stress and cellular senescence in porcine aortic valve interstitial cells
Author: Das, D.
Holmes, A.
Mishra, K.
Rosenkranz, A.
Horowitz, J.
Kennedy, J.
Citation: The Journal of Heart Valve Disease, 2013; 22(5):621-630
Publisher: I C R Publishers
Issue Date: 2013
ISSN: 0966-8519
Statement of
Dividutta Das, Andrew Holmes, Geraldine A. Murphy, Kumaril Mishra, Anke C. Rosenkranz, John D. Horowitz, Jennifer A. Kennedy
Abstract: Background and aim of the study: Aortic valve stenosis is a major cause of valve replacement, particularly in the elderly. TGF-β1 is upregulated in stenotic valves and induces calcification and collagen synthesis in cultured valve interstitial cells. It has been shown previously that TGF-β1 increases reactive oxygen species (ROS) in these cells in association with calcifying nodule formation, but the cellular signaling pathways responsible for these TGF-β1-induced effects are not well defined. Methods: Cultured porcine aortic valve interstitial cells were used to investigate the effects of inhibitors of TGF-β1 signaling pathways on 3H-proline incorporation into the extracellular matrix, the peak number of calcifying nodules formed, redox stress as dichlorofluorescein diacetate (DCF-DA) fluorescence, and senescence-associated β-galactosidase staining. Results: Nodule formation and proline incorporation were inhibited by SB431542, implicating the Smad pathway, by SB203580, implicating the P38 MAPK pathway, and by U0126, implicating the Mek1/2/Erk1/2 pathway in both processes. Fasudil, an inhibitor of the Rho kinase pathway, was selective in inhibiting nodule formation but not proline incorporation. It was verified that Smad 2 phosphorylation, Erk1/2 phosphorylation and p38 MAPK phosphorylation were all induced by TGF-β1, with Smad 2 phosphorylation peaking at 1-2 h and MAPK phosphorylation at 24-48 h. The effect of TGF-β1 on phosphorylation of Smad 2 was inhibited by SB431542, on the phosphorylation of p38 MAPK was inhibited by SB203580, and on the phosphorylation of Erk1/2 was inhibited by U0126. ROS generation in response to TGF-β1, measured as 2,7-dichlorofluorescein-diacetate fluorescence, was inhibited significantly by SB203580 and U0126, implicating both the p38 MAPK and Mek1/2/Erk1/2 signalling pathways. Both pathways also mediated TGF-β1-induced cellular senescence which was localized to cellular aggregates and mature nodules. Conclusion: These data imply that the inhibition of either Smad or MAPK signalling pathways may have a therapeutic benefit in ameliorating the adverse pathological changes associated with aortic valve stenosis.
Keywords: Aortic Valve
Cells, Cultured
Aortic Valve Stenosis
Disease Models, Animal
Mitogen-Activated Protein Kinases
MAP Kinase Signaling System
Transforming Growth Factor beta1
Cellular Senescence
Rights: © ICR Publishers Ltd 2014. All rights reserved
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