The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 Macrophages
Atherosclerosis is developed due to the formation of atheroma plaques in the coronary arteries. In this process, M1 macrophages and vascular smooth muscle cells (VSMCs) are the main functional cells. Inflammatory mediators such as histamine may inflame M1 macrophages. The aim of this study was to de...
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2019-02-01
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| Series: | Iranian Journal of Allergy, Asthma and Immunology |
| Subjects: | |
| Online Access: | https://ijaai.tums.ac.ir/index.php/ijaai/article/view/1943 |
| _version_ | 1818111302965395456 |
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| author | Mohsen Khosravi Mohammad Najafi Abdollah Amirfarhangi Mahdi Karimi Fahimeh Fattahi Mohammad Shabani |
| author_facet | Mohsen Khosravi Mohammad Najafi Abdollah Amirfarhangi Mahdi Karimi Fahimeh Fattahi Mohammad Shabani |
| author_sort | Mohsen Khosravi |
| collection | DOAJ |
| description | Atherosclerosis is developed due to the formation of atheroma plaques in the coronary arteries. In this process, M1 macrophages and vascular smooth muscle cells (VSMCs) are the main functional cells. Inflammatory mediators such as histamine may inflame M1 macrophages. The aim of this study was to determine the effect of M1 macrophage secretion contents on the gene and protein expression levels of focal adhesion kinase (FAK), vasodilator-stimulated phosphoprotein (VASP), and thrombospondin1 (THBS1). Whole blood samples from the six healthy subjects (stenosis<5%), and six patients (stenosis>70%) were prepared and peripheral blood mononuclear cells (PBMCs) were isolated. Then monocytes were differentiated into M1 macrophages using 100 ng/mL granulocyte-macrophage colony stimulating factor (GM-CSF). The differentiated M1 macrophages were treated with histamine (10-6 M), and their secretion contents were harvested and added to the culture medium of VSMCs. The FAK, VASP, and THBS1 gene expression and protein levels were measured using RT-qPCR and western blot techniques in VSMCs, respectively. The FAK and THBS1 gene expression levels significantly increased in VSMCs after adding secretion contents obtained from histamine-treated M1 macrophages (p=0.023 and 0.05, respectively), while significant results were not observed for VASP gene (p=0.45). In converse with the phosphorylated VASP (pVASP) (p<0.34), the phosphorylated FAK (pFAK) and THBS1 protein levels increased in VSMCs (p<0.001). We concluded that in inflammatory conditions, the immune events could affect the macrophages by histamine. The activated macrophages could locally activate signaling pathways via FAK and THBS1 genes that are effective in the proliferation and migration of VSMCs. |
| first_indexed | 2024-12-11T03:00:53Z |
| format | Article |
| id | doaj.art-aaa6dfa10d7248afbc296c53c65d3a1d |
| institution | Directory Open Access Journal |
| issn | 1735-1502 1735-5249 |
| language | English |
| last_indexed | 2024-12-11T03:00:53Z |
| publishDate | 2019-02-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Allergy, Asthma and Immunology |
| spelling | doaj.art-aaa6dfa10d7248afbc296c53c65d3a1d2022-12-22T01:23:03ZengTehran University of Medical SciencesIranian Journal of Allergy, Asthma and Immunology1735-15021735-52492019-02-0118110.18502/ijaai.v18i1.6321943The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 MacrophagesMohsen Khosravi0Mohammad Najafi1Abdollah Amirfarhangi2Mahdi Karimi3Fahimeh Fattahi4Mohammad Shabani5Department of Biochemistry, Iran University of Medical Sciences, Tehran, IranDepartment of Biochemistry, Iran University of Medical Sciences, Tehran, Iran AND Molecular and Cellular Research Center, Iran University of Medical Sciences, Tehran, IranDepartment of Cardiology, Hazrat Rasool Akrsm Hospital, Iran University of Medical Sciences, Tehran, IranDepartment of Medical Nanotechnology, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, IranOncopathology Research Center, Iran University of Medical Sciences, Tehran, Iran AND Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, IranDepartment of Biochemistry, Iran University of Medical Sciences, Tehran, IranAtherosclerosis is developed due to the formation of atheroma plaques in the coronary arteries. In this process, M1 macrophages and vascular smooth muscle cells (VSMCs) are the main functional cells. Inflammatory mediators such as histamine may inflame M1 macrophages. The aim of this study was to determine the effect of M1 macrophage secretion contents on the gene and protein expression levels of focal adhesion kinase (FAK), vasodilator-stimulated phosphoprotein (VASP), and thrombospondin1 (THBS1). Whole blood samples from the six healthy subjects (stenosis<5%), and six patients (stenosis>70%) were prepared and peripheral blood mononuclear cells (PBMCs) were isolated. Then monocytes were differentiated into M1 macrophages using 100 ng/mL granulocyte-macrophage colony stimulating factor (GM-CSF). The differentiated M1 macrophages were treated with histamine (10-6 M), and their secretion contents were harvested and added to the culture medium of VSMCs. The FAK, VASP, and THBS1 gene expression and protein levels were measured using RT-qPCR and western blot techniques in VSMCs, respectively. The FAK and THBS1 gene expression levels significantly increased in VSMCs after adding secretion contents obtained from histamine-treated M1 macrophages (p=0.023 and 0.05, respectively), while significant results were not observed for VASP gene (p=0.45). In converse with the phosphorylated VASP (pVASP) (p<0.34), the phosphorylated FAK (pFAK) and THBS1 protein levels increased in VSMCs (p<0.001). We concluded that in inflammatory conditions, the immune events could affect the macrophages by histamine. The activated macrophages could locally activate signaling pathways via FAK and THBS1 genes that are effective in the proliferation and migration of VSMCs.https://ijaai.tums.ac.ir/index.php/ijaai/article/view/1943AtherosclerosisFocal adhesion kinaseHistamineMacrophagesThrombospondin 1Vascular smooth muscle cells |
| spellingShingle | Mohsen Khosravi Mohammad Najafi Abdollah Amirfarhangi Mahdi Karimi Fahimeh Fattahi Mohammad Shabani The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 Macrophages Iranian Journal of Allergy, Asthma and Immunology Atherosclerosis Focal adhesion kinase Histamine Macrophages Thrombospondin 1 Vascular smooth muscle cells |
| title | The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 Macrophages |
| title_full | The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 Macrophages |
| title_fullStr | The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 Macrophages |
| title_full_unstemmed | The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 Macrophages |
| title_short | The Increase of pFAK and THBS1 Protein and Gene Expression Levels in Vascular Smooth Muscle Cells by Histamine-treated M1 Macrophages |
| title_sort | increase of pfak and thbs1 protein and gene expression levels in vascular smooth muscle cells by histamine treated m1 macrophages |
| topic | Atherosclerosis Focal adhesion kinase Histamine Macrophages Thrombospondin 1 Vascular smooth muscle cells |
| url | https://ijaai.tums.ac.ir/index.php/ijaai/article/view/1943 |
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