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Updated: Aug 13, 2026
Quantitation of Endothelial Cell Adhesiveness In Vitro
Published on: June 18, 2015
Endothelin-1 directly modulates its own secretion: studies utilising the cell immunoblot technique
John J Evans1, Alber H Youssef, Timothy G Yandle
1Department of Obstetrics and Gynaecology, Christchurch School of Medicine and Health Sciences, Christchurch Women's Hospital, Private Bag 4711, Christchurch, New Zealand. john.evans@chmeds.ac.nz
Insights
Endothelin-1 (ET-1) can stimulate its own release from human endothelial cells, a process influenced by angiotensin-II. Receptor antagonists like bosentan and BQ788 block this self-stimulation and basal ET-1 secretion.
Area of Science:
- Cardiovascular Biology
- Endothelial Cell Function
- Peptide Signaling
Background:
- Endothelin-1 (ET-1) is a potent vasoconstrictor crucial for vascular regulation.
- Elevated ET-1 levels are linked to various cardiovascular disorders.
- Mechanisms controlling ET-1 secretion remain incompletely understood.
Purpose of the Study:
- To investigate the autocrine regulation of endothelin-1 secretion by human endothelial cells.
- To determine if ET-1 influences its own release.
- To assess the impact of angiotensin-II and ET-1 receptor antagonists on ET-1 secretion.
Main Methods:
- A cell immunoblot technique was utilized to detect local ET-1 secretion from individual human vascular endothelial cells.
- Immunohistochemical methods were employed to quantify intracellular and secreted ET-1.
- Human endothelial cells were incubated with ET-1, angiotensin-II, bosentan, or BQ788.
Main Results:
- Both endothelin-1 and angiotensin-II significantly increased the number of endothelial cells secreting ET-1.
- Incubation with endothelin receptor antagonists bosentan and BQ788 reduced the number of ET-1 secreting cells.
- Basal ET-1 secretion was inhibited by bosentan and BQ788.
Conclusions:
- Human endothelial cells possess an autocrine pathway where endothelin-1 stimulates its own release.
- Endothelin-1 and angiotensin-II act as positive regulators of ET-1 secretion.
- Bosentan and BQ788 effectively inhibit both basal and stimulated ET-1 secretion, highlighting their therapeutic potential.
Abstract:
Endothelin-1 is an important factor in vasoregulation and circulating levels of the peptide are increased in a number of cardiovascular disorders. However, control of endothelin-1 secretion is only sketchily understood. The possibility that endothelin-1 influences its own release was investigated. A cell immunoblot method, which can detect local secretion of peptide from individual human vascular endothelial cells, was employed. Cells were dispersed onto a protein-binding membrane. Endothelin-1 in cells or secreted and adhering to the protein-binding membrane outside the cells was detected using immunohistochemical techniques. The numbers of cells that contained endothelin-1 and secreted endothelin-1 were counted after the cells had been incubated in control conditions, or with added endothelin-1, angiotensin-II, or endothelin receptor antagonists, bosentan and BQ788. Endothelin-1 and angiotensin-II increased the numbers of cells that secreted endothelin-1. On the other hand, bosentan and BQ788 caused a reduction in the numbers of endothelin-1-secreting cells. These results indicate that human endothelial cells contain a pathway by which endothelin-1 induces its own release. The receptor antagonists, bosentan and BQ788, inhibited basal secretion of endothelin-1.
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