Related Experiment Video
Updated: Aug 8, 2026
Generation of an Immortalized Murine Brain Microvascular Endothelial Cell Line as an In Vitro Blood Brain Barrier Model
Published on: August 29, 2012
Endothelin-1 and monocyte chemoattractant protein-1 modulation in ischemia and human brain-derived endothelial cell
P Chen1, M Shibata, R Zidovetzki
1Department of Pathology, University of Southern California, School of Medicine, Los Angeles, CA 90033, USA.
Abstract:
Brain tissue damage due to ischemia/reperfusion has been shown to be caused, in part, by activated macrophages infiltrating into the post-ischemic brain. Using the Middle Cerebral Artery Occlusion (MCAO) mouse model, this study demonstrated that, in vivo, both endothelin-1 (Et-1), a potent vasoconstrictor, and the macrophage chemokine, monocyte chemoattractant factor-1 (MCP-1) are induced in ischemia. Further studies, using human brain-derived endothelial cells (CNS-EC), showed that in vitro, Et-1 can directly stimulate MCP-1 mRNA expression and MCP-1 protein; and this Et-1-induced MCP-1 production is mediated by the ET(A) receptor. Inflammatory cytokines, tumor necrosis factor alpha and interleukin-1beta, functioned additively and synergistically, respectively, with Et-1 to increase this MCP-1 production. Partial elucidation of the signal transduction pathways involved in Et-1-induced MCP-1 production demonstrated that protein kinase C-, but not cAMP-dependent pathways are involved. These data demonstrate that Et-1, functioning as an inflammatory peptide, increased levels of MCP-1, suggesting a mechanism for chemokine regulation during ischemia/reperfusion injury.
Insights
Endothelin-1 (Et-1) increases monocyte chemoattractant protein-1 (MCP-1) during brain ischemia/reperfusion injury. This suggests a key mechanism involving Et-1 in inflammatory cell recruitment and brain damage.
Area of Science:
- Neuroscience
- Immunology
- Cardiovascular Biology
Background:
- Brain tissue damage from ischemia/reperfusion involves infiltrating macrophages.
- Macrophage recruitment is mediated by chemokines like monocyte chemoattractant protein-1 (MCP-1).
Purpose of the Study:
- To investigate the role of endothelin-1 (Et-1) in regulating MCP-1 during ischemia/reperfusion (I/R) injury.
- To elucidate the signaling pathways involved in Et-1-induced MCP-1 production.
Main Methods:
- Utilized the Middle Cerebral Artery Occlusion (MCAO) mouse model for in vivo studies.
- Employed human brain-derived endothelial cells (CNS-EC) for in vitro experiments.
- Analyzed MCP-1 mRNA and protein expression, and investigated signaling pathways (PKC, cAMP).
Main Results:
- In vivo, both Et-1 and MCP-1 were induced during ischemia in the MCAO model.
- In vitro, Et-1 directly stimulated MCP-1 expression via the ET(A) receptor.
- Inflammatory cytokines tumor necrosis factor alpha and interleukin-1beta enhanced Et-1-induced MCP-1 production.
- Protein kinase C (PKC) pathways, but not cAMP, were involved in Et-1-induced MCP-1 production.
Conclusions:
- Et-1 acts as an inflammatory peptide that upregulates MCP-1 during I/R injury.
- Et-1-mediated MCP-1 induction represents a significant mechanism in chemokine regulation following ischemic stroke.
- Findings suggest potential therapeutic targets for mitigating brain damage in stroke.
