The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases
Information about mechanical strain in the extracellular space is conducted along collagen fibers connected with integrins and then transmitted within cells. An aim of the study is to verify the hypothesis that the stiffness of cardiac human fibroblast substrates exerts a regulatory effect on collag...
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MDPI AG
2021-12-01
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| Online Access: | https://www.mdpi.com/2073-4409/10/12/3506 |
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| author | Małgorzata Gałdyszyńska Paulina Radwańska Jacek Szymański Jacek Drobnik |
| author_facet | Małgorzata Gałdyszyńska Paulina Radwańska Jacek Szymański Jacek Drobnik |
| author_sort | Małgorzata Gałdyszyńska |
| collection | DOAJ |
| description | Information about mechanical strain in the extracellular space is conducted along collagen fibers connected with integrins and then transmitted within cells. An aim of the study is to verify the hypothesis that the stiffness of cardiac human fibroblast substrates exerts a regulatory effect on collagen metabolism via integrin α2β1 and downstream signaling. The experiments were performed on human cardiac fibroblasts cultured on stiff or soft polyacrylamide gels. Extracellular and intracellular collagen content, metalloproteinase-1 (MMP-1), metalloproteinase-9 (MMP-9) and expression of the α1 chain of the procollagen type I gene (<i>Col1A1</i>) were elevated in cultures settled on soft substrate. The substrate stiffness did not modify tissue inhibitors of matrix metalloproteinase capacity (TIMPs 1–4). Integrin α2β1 inhibition (TC-I 15) or α2 subunit silencing resulted in augmentation of collagen content within the culture. Expression of <i>Col1A1</i> and <i>Col3A1</i> genes was increased in TC-I 15-treated fibroblasts. Total and phosphorylated levels of both FAK and Src kinases were elevated in fibroblasts cultured on stiff substrate. Inhibition of FAK (FAK kinase inhibitor 14) or Src kinase (AZM 47527) increased collagen content within the culture. The substrate stiffness exerted a regulatory influence on collagen metabolism via integrin α2β1 and its downstream signaling (FAK and Src kinases) in cardiac fibroblasts. |
| first_indexed | 2024-03-10T04:25:58Z |
| format | Article |
| id | doaj.art-993295e81dc2428ea4c19441686483e4 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T04:25:58Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-993295e81dc2428ea4c19441686483e42023-11-23T07:38:43ZengMDPI AGCells2073-44092021-12-011012350610.3390/cells10123506The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src KinasesMałgorzata Gałdyszyńska0Paulina Radwańska1Jacek Szymański2Jacek Drobnik3Laboratory of Connective Tissue Metabolism, Department of Pathophysiology, Medical University of Lodz, Żeligowskiego 7/9, 90-752 Lodz, PolandLaboratory of Connective Tissue Metabolism, Department of Pathophysiology, Medical University of Lodz, Żeligowskiego 7/9, 90-752 Lodz, PolandCentral Scientific Laboratory, Medical University of Lodz, Mazowiecka 6/8, 92-215 Lodz, PolandLaboratory of Connective Tissue Metabolism, Department of Pathophysiology, Medical University of Lodz, Żeligowskiego 7/9, 90-752 Lodz, PolandInformation about mechanical strain in the extracellular space is conducted along collagen fibers connected with integrins and then transmitted within cells. An aim of the study is to verify the hypothesis that the stiffness of cardiac human fibroblast substrates exerts a regulatory effect on collagen metabolism via integrin α2β1 and downstream signaling. The experiments were performed on human cardiac fibroblasts cultured on stiff or soft polyacrylamide gels. Extracellular and intracellular collagen content, metalloproteinase-1 (MMP-1), metalloproteinase-9 (MMP-9) and expression of the α1 chain of the procollagen type I gene (<i>Col1A1</i>) were elevated in cultures settled on soft substrate. The substrate stiffness did not modify tissue inhibitors of matrix metalloproteinase capacity (TIMPs 1–4). Integrin α2β1 inhibition (TC-I 15) or α2 subunit silencing resulted in augmentation of collagen content within the culture. Expression of <i>Col1A1</i> and <i>Col3A1</i> genes was increased in TC-I 15-treated fibroblasts. Total and phosphorylated levels of both FAK and Src kinases were elevated in fibroblasts cultured on stiff substrate. Inhibition of FAK (FAK kinase inhibitor 14) or Src kinase (AZM 47527) increased collagen content within the culture. The substrate stiffness exerted a regulatory influence on collagen metabolism via integrin α2β1 and its downstream signaling (FAK and Src kinases) in cardiac fibroblasts.https://www.mdpi.com/2073-4409/10/12/3506collagencell culturecardiac fibroblastsfocal adhesion kinaseheartintegrin α2β1 |
| spellingShingle | Małgorzata Gałdyszyńska Paulina Radwańska Jacek Szymański Jacek Drobnik The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases Cells collagen cell culture cardiac fibroblasts focal adhesion kinase heart integrin α2β1 |
| title | The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases |
| title_full | The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases |
| title_fullStr | The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases |
| title_full_unstemmed | The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases |
| title_short | The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases |
| title_sort | stiffness of cardiac fibroblast substrates exerts a regulatory influence on collagen metabolism via α2β1 integrin fak and src kinases |
| topic | collagen cell culture cardiac fibroblasts focal adhesion kinase heart integrin α2β1 |
| url | https://www.mdpi.com/2073-4409/10/12/3506 |
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