Glycogen synthase kinase-3 (GSK-3) regulates TGF-β1-induced differentiation of pulmonary fibroblasts
| Autor: | Baarsma, Hoeke A, Engelbertink, Lilian HJM, van Hees, Lonneke J, Menzen, Mark H, Meurs, Herman, Timens, Wim, Postma, Dirkje S, Kerstjens, Huib AM, Gosens, Reinoud |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Cell Differentiation
Fibroblasts Research Papers Recombinant Proteins Cell Line Transforming Growth Factor beta1 Glycogen Synthase Kinase 3 Pulmonary Disease Chronic Obstructive Gene Expression Regulation Humans Gene Silencing RNA Messenger Phosphorylation Cyclic AMP Response Element-Binding Protein Myofibroblasts Lung Protein Kinase Inhibitors Protein Processing Post-Translational Biomarkers Cells Cultured Signal Transduction |
| Popis: | Chronic lung diseases such as asthma, COPD and pulmonary fibrosis are characterized by abnormal extracellular matrix (ECM) turnover. TGF-β is a key mediator stimulating ECM production by recruiting and activating lung fibroblasts and initiating their differentiation process into more active myofibroblasts. Glycogen synthase kinase-3 (GSK-3) regulates various intracellular signalling pathways; its role in TGF-β₁-induced myofibroblast differentiation is currently largely unknown.To determine the contribution of GSK-3 signalling in TGF-β₁-induced myofibroblast differentiation.We used MRC5 human lung fibroblasts and primary pulmonary fibroblasts of individuals with and without COPD. Protein and mRNA expression were determined by immunoblotting and RT-PCR analysis respectively.Stimulation of MRC5 and primary human lung fibroblasts with TGF-β₁ resulted in time- and dose-dependent increases of α-sm-actin and fibronectin expression, indicative of myofibroblast differentiation. Pharmacological inhibition of GSK-3 by SB216763 dose-dependently attenuated TGF-β₁-induced expression of these myofibroblasts markers. Moreover, silencing of GSK-3 by siRNA or pharmacological inhibition by CT/CHIR99021 fully inhibited the TGF-β₁-induced expression of α-sm-actin and fibronectin. The effect of GSK-3 inhibition on α-sm-actin expression was similar in fibroblasts from individuals with and without COPD. Neither smad, NF-κB nor ERK1/2 were involved in the inhibitory actions of GSK-3 inhibition by SB126763 on myofibroblast differentiation. Rather, SB216763 increased the phosphorylation of CREB, which in its phosphorylated form acts as a functional antagonist of TGF-β/smad signalling.We demonstrate that GSK-3 signalling regulates TGF-β₁-induced myofibroblast differentiation by regulating CREB phosphorylation. GSK-3 may constitute a useful target for treatment of chronic lung diseases. |
| Databáze: | OpenAIRE |
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