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Comprehensive transcriptome of proteases and protease inhibitors in vascular cells
Guo-Ping Shi1, Gregory M Dolganov
1Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA. gshi@rics.bwh. harvard.edu
Insights
Vascular cells express various proteolytic enzymes and inhibitors, crucial for maintaining vascular integrity. This study maps these protease profiles in smooth muscle cells, endothelial cells, and macrophages under inflammatory conditions, aiding research into vascular diseases.
Area of Science:
- Vascular Biology
- Molecular Biology
- Proteomics
Background:
- Vascular cells, including smooth muscle cells, endothelial cells, and macrophages, are vital for vascular wall integrity.
- Inflammation and altered gene expression, particularly of proteolytic enzymes, are implicated in vascular diseases like atherosclerosis and cerebral ischemia.
Purpose of the Study:
- To investigate the mRNA expression profiles of proteolytic enzymes and their inhibitors in human vascular cells.
- To understand how common proinflammatory mediators influence protease expression in these cells, relevant to vascular disease pathogenesis.
Main Methods:
- Primary human saphenous vein smooth muscle cells, endothelial cells, and monocyte-derived macrophages were isolated from three donors.
- Cells were stimulated with five key proinflammatory mediators: interleukin-1beta, interferon-gamma, tumor necrosis factor-alpha, basic fibroblast growth factor, and vascular endothelial growth factor.
- Quantitative real-time PCR was employed to analyze the mRNA levels of 49 selected proteolytic enzymes and inhibitors from four protease families.
Main Results:
- The study identified distinct expression patterns of proteolytic enzymes and their inhibitors across different vascular cell types.
- Proinflammatory cytokine stimulation led to significant alterations in the expression of these proteases, highlighting their role in inflammatory responses.
- A comprehensive dataset of vascular cell protease expression profiles under inflammatory conditions was generated.
Conclusions:
- The findings provide a broad overview of vascular cell protease expression under inflammation, essential for understanding vascular tissue remodeling.
- While donor variability exists, the data offers critical insights for research into inflammation-associated vascular diseases.
- This study establishes a foundation for further investigations into the specific roles of these proteases in vascular health and disease.
Background And Purpose:
Smooth muscle cells, endothelial cells, and macrophages are essential components of the vasculature, of which the homeostatic gene expression participate importantly in the maintenance of vascular wall integrity. The pathogenesis of vascular diseases, such as cerebral ischemia, atherosclerosis, and abdominal aortic aneurysms, often associates with inflammation and altered gene expression, including proteolytic enzymes that play multiple and important roles in extracellular matrix degradation, cell proliferation and migration, and latent enzyme or growth factor activation.
Methods:
Human saphenous vein smooth muscle cells, endothelial cells, and monocyte-derived macrophages from 3 independent donors were stimulated with interleukin 1beta, interferon gamma, tumor necrosis factor alpha, basic fibroblast growth factor, and vascular endothelial growth factor, 5 common proinflammatory mediators often found in diseased human microvessels and macrovessels. Quantitative real-time PCR was used to examine the mRNA levels of 49 proteolytic enzymes and their inhibitors, selected from 4 protease families, in these vascular cells.
Conclusions:
Although primary cultured cells from different donors may behave differently in response to these proinflammatory cytokines, data from this study revealed a broad view of vascular cell protease expression profiles under inflammatory conditions, critical to studies of inflammation-associated vascular tissue remodeling.
