Semicarbazide-Sensitive Amine Oxidase (SSAO) and Lysyl Oxidase (LOX) Association in Rat Aortic Vascular Smooth Muscle Cells
Vascular smooth muscle cells (VSMCs) are the main stromal cells in the medial layer of the vascular wall. These cells produce the extracellular matrix (ECM) and are involved in many pathological changes in the vascular wall. Semicarbazide-sensitive amine oxidase (SSAO) and lysyl oxidase (LOX) are va...
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MDPI AG
2022-10-01
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| Series: | Biomolecules |
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| author | Vesna Manasieva Shori Thakur Lisa A. Lione Jessal Patel Anwar Baydoun John Skamarauskas |
| author_facet | Vesna Manasieva Shori Thakur Lisa A. Lione Jessal Patel Anwar Baydoun John Skamarauskas |
| author_sort | Vesna Manasieva |
| collection | DOAJ |
| description | Vascular smooth muscle cells (VSMCs) are the main stromal cells in the medial layer of the vascular wall. These cells produce the extracellular matrix (ECM) and are involved in many pathological changes in the vascular wall. Semicarbazide-sensitive amine oxidase (SSAO) and lysyl oxidase (LOX) are vascular enzymes associated with the development of atherosclerosis. In the vascular smooth muscle cells, increased SSAO activity elevates reactive oxygen species (ROS) and induces VSMCs death; increased LOX induces chemotaxis through hydrogen peroxide dependent mechanisms; and decreased LOX contributes to endothelial dysfunction. This study investigates the relationship between SSAO and LOX in VSMCs by studying their activity, protein, and mRNA levels during VSMCs passaging and after silencing the LOX gene, while using their respective substrates and inhibitors. At the basal level, LOX activity decreased with passage and its protein expression was maintained between passages. βAPN abolished LOX activity (** <i>p</i> < 0.01 for 8 vs. 3 and * <i>p</i> < 0.05 for 5 vs. 8) and had no effect on LOX protein and mRNA levels. MDL72527 reduced LOX activity at passage 3 and 5 (<sup>##</sup> <i>p</i> < 0.01) and had no effect on LOX protein, and mRNA expression. At the basal level, SSAO activity also decreased with passage, and its protein expression was maintained between passages. MDL72527 abolished SSAO activity (**** <i>p</i> < 0.0001 for 8 vs. 3 and * <i>p</i> < 0.05 for 5 vs. 8), VAP-1 expression at passage 5 (** <i>p</i> < 0.01) and 8 (**** <i>p</i> < 0.0001), and Aoc3 mRNA levels at passage 8 (* <i>p</i> < 0.05). βAPN inhibited SSAO activity (**** <i>p</i> < 0.0001 for 5 vs. 3 and 8 vs. 3 and * <i>p</i> < 0.05 for 5 vs. 8), VAP-1 expression at passage 3 (* <i>p</i> < 0.05), and Aoc3 mRNA levels at passage 3 (* <i>p</i> < 0.05). Knockdown of the LOX gene (**** <i>p</i> < 0.0001 for Si6 vs. Sictrl and *** <i>p</i> < 0.001 for Si8 vs. Sictrl) and LOX protein (** <i>p</i> < 0.01 for Si6 and Si8 vs. Sictrl) in VSMCs at passage 3 resulted in a reduction in Aoc3 mRNA (<sup>####</sup> <i>p</i> < 0.0001 for Si6 vs. Sictrl and <sup>###</sup> <i>p</i> < 0.001 for Si8 vs. Sictrl) and VAP-1 protein (<sup>#</sup> <i>p</i> < 0.05 for Si8 vs. Sictrl). These novel findings demonstrate a passage dependent decrease in LOX activity and increase in SSAO activity in rat aortic VSMCs and show an association between both enzymes in early passage rat aortic VSMCs, where LOX was identified as a regulator of SSAO activity, protein, and mRNA expression. |
| first_indexed | 2024-03-09T19:14:33Z |
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| language | English |
| last_indexed | 2024-03-09T19:14:33Z |
| publishDate | 2022-10-01 |
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| series | Biomolecules |
| spelling | doaj.art-c862ab248cf44c07b9808abe8d60a3fc2023-11-24T03:52:41ZengMDPI AGBiomolecules2218-273X2022-10-011211156310.3390/biom12111563Semicarbazide-Sensitive Amine Oxidase (SSAO) and Lysyl Oxidase (LOX) Association in Rat Aortic Vascular Smooth Muscle CellsVesna Manasieva0Shori Thakur1Lisa A. Lione2Jessal Patel3Anwar Baydoun4John Skamarauskas5Imperial Biomedical Research Centre, Hammersmith Hospital, Department of Metabolism, Digestion and Reproduction, School of Medicine, Imperial College, London W12 0NN, UKCentre for Health Services and Clinical Research, Basic and Clinical Science Unit, Department of Clinical, Pharmaceutical and Biological Science, School of Life and Medical Sciences, University of Hertfordshire, Hatfield AL10 9AB, UKCentre for Health Services and Clinical Research, Basic and Clinical Science Unit, Department of Clinical, Pharmaceutical and Biological Science, School of Life and Medical Sciences, University of Hertfordshire, Hatfield AL10 9AB, UKCentre for Health Services and Clinical Research, Basic and Clinical Science Unit, Department of Clinical, Pharmaceutical and Biological Science, School of Life and Medical Sciences, University of Hertfordshire, Hatfield AL10 9AB, UKLeicester Institute of Pharmaceutical Innovation for Integrated Care (LIPIIC), Health and Life Sciences, De Montfort University, The Gateway, Leicester LE1 9BH, UKCentre for Health Services and Clinical Research, Basic and Clinical Science Unit, Department of Clinical, Pharmaceutical and Biological Science, School of Life and Medical Sciences, University of Hertfordshire, Hatfield AL10 9AB, UKVascular smooth muscle cells (VSMCs) are the main stromal cells in the medial layer of the vascular wall. These cells produce the extracellular matrix (ECM) and are involved in many pathological changes in the vascular wall. Semicarbazide-sensitive amine oxidase (SSAO) and lysyl oxidase (LOX) are vascular enzymes associated with the development of atherosclerosis. In the vascular smooth muscle cells, increased SSAO activity elevates reactive oxygen species (ROS) and induces VSMCs death; increased LOX induces chemotaxis through hydrogen peroxide dependent mechanisms; and decreased LOX contributes to endothelial dysfunction. This study investigates the relationship between SSAO and LOX in VSMCs by studying their activity, protein, and mRNA levels during VSMCs passaging and after silencing the LOX gene, while using their respective substrates and inhibitors. At the basal level, LOX activity decreased with passage and its protein expression was maintained between passages. βAPN abolished LOX activity (** <i>p</i> < 0.01 for 8 vs. 3 and * <i>p</i> < 0.05 for 5 vs. 8) and had no effect on LOX protein and mRNA levels. MDL72527 reduced LOX activity at passage 3 and 5 (<sup>##</sup> <i>p</i> < 0.01) and had no effect on LOX protein, and mRNA expression. At the basal level, SSAO activity also decreased with passage, and its protein expression was maintained between passages. MDL72527 abolished SSAO activity (**** <i>p</i> < 0.0001 for 8 vs. 3 and * <i>p</i> < 0.05 for 5 vs. 8), VAP-1 expression at passage 5 (** <i>p</i> < 0.01) and 8 (**** <i>p</i> < 0.0001), and Aoc3 mRNA levels at passage 8 (* <i>p</i> < 0.05). βAPN inhibited SSAO activity (**** <i>p</i> < 0.0001 for 5 vs. 3 and 8 vs. 3 and * <i>p</i> < 0.05 for 5 vs. 8), VAP-1 expression at passage 3 (* <i>p</i> < 0.05), and Aoc3 mRNA levels at passage 3 (* <i>p</i> < 0.05). Knockdown of the LOX gene (**** <i>p</i> < 0.0001 for Si6 vs. Sictrl and *** <i>p</i> < 0.001 for Si8 vs. Sictrl) and LOX protein (** <i>p</i> < 0.01 for Si6 and Si8 vs. Sictrl) in VSMCs at passage 3 resulted in a reduction in Aoc3 mRNA (<sup>####</sup> <i>p</i> < 0.0001 for Si6 vs. Sictrl and <sup>###</sup> <i>p</i> < 0.001 for Si8 vs. Sictrl) and VAP-1 protein (<sup>#</sup> <i>p</i> < 0.05 for Si8 vs. Sictrl). These novel findings demonstrate a passage dependent decrease in LOX activity and increase in SSAO activity in rat aortic VSMCs and show an association between both enzymes in early passage rat aortic VSMCs, where LOX was identified as a regulator of SSAO activity, protein, and mRNA expression.https://www.mdpi.com/2218-273X/12/11/1563semicarbazide sensitive amine oxidase (SSAO)lysyl oxidase (LOX)copper amine oxidasesvascular smooth muscle cellsMDL72527βAPN |
| spellingShingle | Vesna Manasieva Shori Thakur Lisa A. Lione Jessal Patel Anwar Baydoun John Skamarauskas Semicarbazide-Sensitive Amine Oxidase (SSAO) and Lysyl Oxidase (LOX) Association in Rat Aortic Vascular Smooth Muscle Cells Biomolecules semicarbazide sensitive amine oxidase (SSAO) lysyl oxidase (LOX) copper amine oxidases vascular smooth muscle cells MDL72527 βAPN |
| title | Semicarbazide-Sensitive Amine Oxidase (SSAO) and Lysyl Oxidase (LOX) Association in Rat Aortic Vascular Smooth Muscle Cells |
| title_full | Semicarbazide-Sensitive Amine Oxidase (SSAO) and Lysyl Oxidase (LOX) Association in Rat Aortic Vascular Smooth Muscle Cells |
| title_fullStr | Semicarbazide-Sensitive Amine Oxidase (SSAO) and Lysyl Oxidase (LOX) Association in Rat Aortic Vascular Smooth Muscle Cells |
| title_full_unstemmed | Semicarbazide-Sensitive Amine Oxidase (SSAO) and Lysyl Oxidase (LOX) Association in Rat Aortic Vascular Smooth Muscle Cells |
| title_short | Semicarbazide-Sensitive Amine Oxidase (SSAO) and Lysyl Oxidase (LOX) Association in Rat Aortic Vascular Smooth Muscle Cells |
| title_sort | semicarbazide sensitive amine oxidase ssao and lysyl oxidase lox association in rat aortic vascular smooth muscle cells |
| topic | semicarbazide sensitive amine oxidase (SSAO) lysyl oxidase (LOX) copper amine oxidases vascular smooth muscle cells MDL72527 βAPN |
| url | https://www.mdpi.com/2218-273X/12/11/1563 |
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