GSK-3β in mouse fibroblasts controls wound healing and fibrosis through an endothelin-1–dependent mechanism
Mohit Kapoor, … , David J. Abraham, Andrew Leask
Mohit Kapoor, … , David J. Abraham, Andrew Leask
Published September 18, 2008
Citation Information: J Clin Invest. 2008;118(10):3279-3290. https://doi.org/10.1172/JCI35381.
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Research Article Dermatology

GSK-3β in mouse fibroblasts controls wound healing and fibrosis through an endothelin-1–dependent mechanism

Abstract

Glycogen synthase kinase–3 (GSK-3) is a widely expressed and highly conserved serine/threonine protein kinase encoded by 2 genes, GSK3A and GSK3B. GSK-3 is thought to be involved in tissue repair and fibrogenesis, but its role in these processes is currently unknown. To investigate the function of GSK-3β in fibroblasts, we generated mice harboring a fibroblast-specific deletion of Gsk3b and evaluated their wound-healing and fibrogenic responses. We have shown that Gsk3b-conditional-KO mice (Gsk3b-CKO mice) exhibited accelerated wound closure, increased fibrogenesis, and excessive scarring compared with control mice. In addition, Gsk3b-CKO mice showed elevated collagen production, decreased cell apoptosis, elevated levels of profibrotic α-SMA, and increased myofibroblast formation during wound healing. In cultured Gsk3b-CKO fibroblasts, adhesion, spreading, migration, and contraction were enhanced. Both Gsk3b-CKO mice and fibroblasts showed elevated expression and production of endothelin-1 (ET-1) compared with control mice and cells. Antagonizing ET-1 reversed the phenotype of Gsk3b-CKO fibroblasts and mice. Thus, GSK-3β appears to control the progression of wound healing and fibrosis by modulating ET-1 levels. These results suggest that targeting the GSK-3β pathway or ET-1 may be of benefit in controlling tissue repair and fibrogenic responses in vivo.

Authors

Mohit Kapoor, Shangxi Liu, Xu Shi-wen, Kun Huh, Matthew McCann, Christopher P. Denton, James R. Woodgett, David J. Abraham, Andrew Leask

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Figure 9

The dual ETA/B receptor antagonist bosentan alleviates the phenotype of GSK-3β–deficient mice (in vivo).

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The dual ETA/B receptor antagonist bosentan alleviates the phenotype of ...
(A and B) Kinetics of wound closure was determined in Gsk3b-C and Gsk3b-CKO mice treated with bosentan (100 mg/kg) or gum arabic (vehicle) on days 0, 7, and 10 after wounding. Bosentan treatment significantly (P < 0.05) reversed the enhanced rate of wound closure in Gsk3b-CKO mice without affecting the wound closure of Gsk3b-C mice (n = 4 wounds from 4 mice). Scale bar: 1 mm. (C) Effect of bosentan treatment on collagen synthesis was assessed using van Gieson staining in day 21 wounds. Note the reduction in the intensity of pink/red staining in Gsk3b-CKO animals in the presence of bosentan. Representative data from n = 4 wounds from 4 mice are shown. Scale bar: 100 μm. (D) Hydroxyproline levels. Bosentan (P < 0.05) reversed elevated wound collagen (hydroxyproline) synthesis in Gsk3b-CKO mice on day 21 after wounding (n = 4 wounds from 4 mice). (E) Bosentan (P < 0.05) reduced and reversed elevated α-SMA expression in Gsk3b-CKO mice on day 21 after wounding (n = 4 wounds from 4 mice). Scale bar: 100 μm. (F) Bosentan significantly (P < 0.05) reduced and reversed elevated α-SMA protein expression in Gsk3b-CKO mice on day 21 after wounding (n = 3). *P < 0.05. (G and H) Bosentan (P < 0.05) rescued loss of apoptosis in Gsk3b-CKO mice on day 7 after wounding (n = 4 wounds from 4 mice). Arrows indicate apoptotic cells. Scale bar: 100 μm. *P < 0.05.