Biglycan Involvement in Heart Fibrosis: Modulation of Adenosine 2A Receptor Improves Damage in Immortalized Cardiac Fibroblasts
Cardiac fibrosis is a common pathological feature of different cardiovascular diseases, characterized by the aberrant deposition of extracellular matrix (ECM) proteins in the cardiac interstitium, myofibroblast differentiation and increased fibrillar collagen deposition stimulated by transforming gr...
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MDPI AG
2023-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/2/1784 |
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| author | Michele Scuruchi Federica Mannino Chiara Imbesi Giovanni Pallio Giovanna Vermiglio Gianluca Bagnato Letteria Minutoli Alessandra Bitto Francesco Squadrito Natasha Irrera |
| author_facet | Michele Scuruchi Federica Mannino Chiara Imbesi Giovanni Pallio Giovanna Vermiglio Gianluca Bagnato Letteria Minutoli Alessandra Bitto Francesco Squadrito Natasha Irrera |
| author_sort | Michele Scuruchi |
| collection | DOAJ |
| description | Cardiac fibrosis is a common pathological feature of different cardiovascular diseases, characterized by the aberrant deposition of extracellular matrix (ECM) proteins in the cardiac interstitium, myofibroblast differentiation and increased fibrillar collagen deposition stimulated by transforming growth factor (TGF)-β activation. Biglycan (BGN), a small leucine-rich proteoglycan (SLRPG) integrated within the ECM, plays a key role in matrix assembly and the phenotypic control of cardiac fibroblasts. Moreover, BGN is critically involved in pathological cardiac remodeling through TGF-β binding, thus causing myofibroblast differentiation and proliferation. Adenosine receptors (ARs), and in particular A<sub>2AR</sub>, may play a key role in stimulating fibrotic damage through collagen production/deposition, as a consequence of cyclic AMP (cAMP) and AKT activation. For this reason, A<sub>2AR</sub> modulation could be a useful tool to manage cardiac fibrosis in order to reduce fibrotic scar deposition in heart tissue. Therefore, the aim of the present study was to investigate the possible crosstalk between A<sub>2AR</sub> and BGN modulation in an in vitro model of TGF-β-induced fibrosis. Immortalized human cardiac fibroblasts (IM-HCF) were stimulated with TGF-β at the concentration of 10 ng/mL for 24 h to induce a fibrotic phenotype. After applying the TGF-β stimulus, cells were treated with two different A<sub>2AR</sub> antagonists, Istradefylline and ZM241385, for an additional 24 h, at the concentration of 10 µM and 1 µM, respectively. Both A<sub>2AR</sub> antagonists were able to regulate the oxidative stress induced by TGF-β through intracellular reactive oxygen species (ROS) reduction in IM-HCFs. Moreover, collagen1a1, MMPs 3/9, BGN, caspase-1 and IL-1β gene expression was markedly decreased following A<sub>2AR</sub> antagonist treatment in TGF-β-challenged human fibroblasts. The results obtained for collagen1a1, SMAD3, α-SMA and BGN were also confirmed when protein expression was evaluated; phospho-Akt protein levels were also reduced following Istradefylline and ZM241385 use, thus suggesting that collagen production involves AKT recruited by the A<sub>2AR</sub>. These results suggest that A<sub>2AR</sub> modulation might be an effective therapeutic option to reduce the fibrotic processes involved in heart pathological remodeling. |
| first_indexed | 2024-03-09T12:16:41Z |
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| language | English |
| last_indexed | 2024-03-09T12:16:41Z |
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| spelling | doaj.art-47186d2d4af248e0a7550432968e51ef2023-11-30T22:45:34ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-01-01242178410.3390/ijms24021784Biglycan Involvement in Heart Fibrosis: Modulation of Adenosine 2A Receptor Improves Damage in Immortalized Cardiac FibroblastsMichele Scuruchi0Federica Mannino1Chiara Imbesi2Giovanni Pallio3Giovanna Vermiglio4Gianluca Bagnato5Letteria Minutoli6Alessandra Bitto7Francesco Squadrito8Natasha Irrera9Department of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Biomedical, Dental, Morphological and Functional Imaging Sciences, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Via C. Valeria, 98125 Messina, ItalyCardiac fibrosis is a common pathological feature of different cardiovascular diseases, characterized by the aberrant deposition of extracellular matrix (ECM) proteins in the cardiac interstitium, myofibroblast differentiation and increased fibrillar collagen deposition stimulated by transforming growth factor (TGF)-β activation. Biglycan (BGN), a small leucine-rich proteoglycan (SLRPG) integrated within the ECM, plays a key role in matrix assembly and the phenotypic control of cardiac fibroblasts. Moreover, BGN is critically involved in pathological cardiac remodeling through TGF-β binding, thus causing myofibroblast differentiation and proliferation. Adenosine receptors (ARs), and in particular A<sub>2AR</sub>, may play a key role in stimulating fibrotic damage through collagen production/deposition, as a consequence of cyclic AMP (cAMP) and AKT activation. For this reason, A<sub>2AR</sub> modulation could be a useful tool to manage cardiac fibrosis in order to reduce fibrotic scar deposition in heart tissue. Therefore, the aim of the present study was to investigate the possible crosstalk between A<sub>2AR</sub> and BGN modulation in an in vitro model of TGF-β-induced fibrosis. Immortalized human cardiac fibroblasts (IM-HCF) were stimulated with TGF-β at the concentration of 10 ng/mL for 24 h to induce a fibrotic phenotype. After applying the TGF-β stimulus, cells were treated with two different A<sub>2AR</sub> antagonists, Istradefylline and ZM241385, for an additional 24 h, at the concentration of 10 µM and 1 µM, respectively. Both A<sub>2AR</sub> antagonists were able to regulate the oxidative stress induced by TGF-β through intracellular reactive oxygen species (ROS) reduction in IM-HCFs. Moreover, collagen1a1, MMPs 3/9, BGN, caspase-1 and IL-1β gene expression was markedly decreased following A<sub>2AR</sub> antagonist treatment in TGF-β-challenged human fibroblasts. The results obtained for collagen1a1, SMAD3, α-SMA and BGN were also confirmed when protein expression was evaluated; phospho-Akt protein levels were also reduced following Istradefylline and ZM241385 use, thus suggesting that collagen production involves AKT recruited by the A<sub>2AR</sub>. These results suggest that A<sub>2AR</sub> modulation might be an effective therapeutic option to reduce the fibrotic processes involved in heart pathological remodeling.https://www.mdpi.com/1422-0067/24/2/1784cardiac fibrosisA<sub>2A</sub> receptorbiglycancollagen |
| spellingShingle | Michele Scuruchi Federica Mannino Chiara Imbesi Giovanni Pallio Giovanna Vermiglio Gianluca Bagnato Letteria Minutoli Alessandra Bitto Francesco Squadrito Natasha Irrera Biglycan Involvement in Heart Fibrosis: Modulation of Adenosine 2A Receptor Improves Damage in Immortalized Cardiac Fibroblasts International Journal of Molecular Sciences cardiac fibrosis A<sub>2A</sub> receptor biglycan collagen |
| title | Biglycan Involvement in Heart Fibrosis: Modulation of Adenosine 2A Receptor Improves Damage in Immortalized Cardiac Fibroblasts |
| title_full | Biglycan Involvement in Heart Fibrosis: Modulation of Adenosine 2A Receptor Improves Damage in Immortalized Cardiac Fibroblasts |
| title_fullStr | Biglycan Involvement in Heart Fibrosis: Modulation of Adenosine 2A Receptor Improves Damage in Immortalized Cardiac Fibroblasts |
| title_full_unstemmed | Biglycan Involvement in Heart Fibrosis: Modulation of Adenosine 2A Receptor Improves Damage in Immortalized Cardiac Fibroblasts |
| title_short | Biglycan Involvement in Heart Fibrosis: Modulation of Adenosine 2A Receptor Improves Damage in Immortalized Cardiac Fibroblasts |
| title_sort | biglycan involvement in heart fibrosis modulation of adenosine 2a receptor improves damage in immortalized cardiac fibroblasts |
| topic | cardiac fibrosis A<sub>2A</sub> receptor biglycan collagen |
| url | https://www.mdpi.com/1422-0067/24/2/1784 |
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