The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cells
<h4>Background</h4> The Rho-kinase ROCK II plays a major role in the activation of hepatic stellate cells (HSC), which are the key profibrotic and contractile cells contributing to the development of chronic liver disease. Inhibition of ROCK II ultimately blocks the phosphorylation of th...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2023-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9888688/?tool=EBI |
| _version_ | 1811173566530453504 |
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| author | Nadine Bachtler Sandra Torres Cristina Ortiz Robert Schierwagen Olaf Tyc Christoph Hieber Marie-Luise Berres Caroline Meier Nico Kraus Stefan Zeuzem Bart Nijmeijer Sebas Pronk Jonel Trebicka Sabine Klein |
| author_facet | Nadine Bachtler Sandra Torres Cristina Ortiz Robert Schierwagen Olaf Tyc Christoph Hieber Marie-Luise Berres Caroline Meier Nico Kraus Stefan Zeuzem Bart Nijmeijer Sebas Pronk Jonel Trebicka Sabine Klein |
| author_sort | Nadine Bachtler |
| collection | DOAJ |
| description | <h4>Background</h4> The Rho-kinase ROCK II plays a major role in the activation of hepatic stellate cells (HSC), which are the key profibrotic and contractile cells contributing to the development of chronic liver disease. Inhibition of ROCK II ultimately blocks the phosphorylation of the myosin light chain (MLC) and thus inhibits stress fibre assembly and cell contraction. We investigated the effects of the ROCK inhibitors Y-33075 as well as Y-27632 in murine and human hepatic stellate cells. <h4>Methods</h4> Primary isolated HSC from FVB/NJ mice and the immortalized human HSC line TWNT-4 were culture-activated and incubated with Y-27632 and Y-33075 (10nM to 10μM) for 24h. Protein expression levels were analyzed by Western Blots and transcriptional levels of pro-fibrotic markers and proliferative markers were evaluated using real-time qPCR. Migration was investigated by wound-healing assay. Proliferation was assessed by BrdU assay. Contraction of HSC was measured using 3D collagen matrices after incubation with Y-27632 or Y-33075 in different doses. <h4>Results</h4> Both Rho-kinase inhibitors, Y-27632 and Y-33075, reduced contraction, fibrogenesis and proliferation in activated primary mouse HSC (FVB/NJ) and human HSC line (TWNT-4) significantly. Y-33075 demonstrated a 10-times increased potency compared to Y-27632. Surprisingly, both inhibitors mediated a substantial and unexpected increase in migration of HSC in FVB/NJ. <h4>Conclusion</h4> ROCK inhibition by the tested compounds decreased contraction but increased migration. Y-33075 proved more potent than Y27632 in the inhibition of contraction of HSCs and should be further evaluated in chronic liver disease. |
| first_indexed | 2024-04-10T17:49:04Z |
| format | Article |
| id | doaj.art-ca41659174874eda92b1d6015a48f512 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-10T17:49:04Z |
| publishDate | 2023-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-ca41659174874eda92b1d6015a48f5122023-02-02T22:58:20ZengPublic Library of Science (PLoS)PLoS ONE1932-62032023-01-01181The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cellsNadine BachtlerSandra TorresCristina OrtizRobert SchierwagenOlaf TycChristoph HieberMarie-Luise BerresCaroline MeierNico KrausStefan ZeuzemBart NijmeijerSebas PronkJonel TrebickaSabine Klein<h4>Background</h4> The Rho-kinase ROCK II plays a major role in the activation of hepatic stellate cells (HSC), which are the key profibrotic and contractile cells contributing to the development of chronic liver disease. Inhibition of ROCK II ultimately blocks the phosphorylation of the myosin light chain (MLC) and thus inhibits stress fibre assembly and cell contraction. We investigated the effects of the ROCK inhibitors Y-33075 as well as Y-27632 in murine and human hepatic stellate cells. <h4>Methods</h4> Primary isolated HSC from FVB/NJ mice and the immortalized human HSC line TWNT-4 were culture-activated and incubated with Y-27632 and Y-33075 (10nM to 10μM) for 24h. Protein expression levels were analyzed by Western Blots and transcriptional levels of pro-fibrotic markers and proliferative markers were evaluated using real-time qPCR. Migration was investigated by wound-healing assay. Proliferation was assessed by BrdU assay. Contraction of HSC was measured using 3D collagen matrices after incubation with Y-27632 or Y-33075 in different doses. <h4>Results</h4> Both Rho-kinase inhibitors, Y-27632 and Y-33075, reduced contraction, fibrogenesis and proliferation in activated primary mouse HSC (FVB/NJ) and human HSC line (TWNT-4) significantly. Y-33075 demonstrated a 10-times increased potency compared to Y-27632. Surprisingly, both inhibitors mediated a substantial and unexpected increase in migration of HSC in FVB/NJ. <h4>Conclusion</h4> ROCK inhibition by the tested compounds decreased contraction but increased migration. Y-33075 proved more potent than Y27632 in the inhibition of contraction of HSCs and should be further evaluated in chronic liver disease.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9888688/?tool=EBI |
| spellingShingle | Nadine Bachtler Sandra Torres Cristina Ortiz Robert Schierwagen Olaf Tyc Christoph Hieber Marie-Luise Berres Caroline Meier Nico Kraus Stefan Zeuzem Bart Nijmeijer Sebas Pronk Jonel Trebicka Sabine Klein The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cells PLoS ONE |
| title | The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cells |
| title_full | The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cells |
| title_fullStr | The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cells |
| title_full_unstemmed | The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cells |
| title_short | The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cells |
| title_sort | non selective rho kinase inhibitors y 27632 and y 33075 decrease contraction but increase migration in murine and human hepatic stellate cells |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9888688/?tool=EBI |
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