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Perilla ketone increases endothelial cell monolayer permeability in vitro
C M Waters1, J S Alexander, T R Harris
1Department of Biomedical Engineering, Vanderbilt University, Nashville, Tennessee 37235.
Summary
Perilla ketone (PK) rapidly and reversibly increases endothelial permeability, a key factor in lung toxin effects. This occurs directly, without needing cytochrome P-450 activation, impacting actin microfilaments.
Area of Science:
- Toxicology
- Cell Biology
- Biochemistry
Background:
- Perilla ketone (PK) is a lung toxin causing pulmonary edema in animals.
- The precise mechanism of PK toxicity remains unknown.
- Understanding PK's cellular effects is crucial for animal health and toxicology.
Purpose of the Study:
- To investigate if Perilla ketone (PK) directly impacts endothelial cell permeability.
- To elucidate the cellular mechanisms underlying PK-induced toxicity.
- To determine the role of cytochrome P-450 in PK activation.
Main Methods:
- Bovine aortic endothelial cells cultured on microcarrier beads in a chromatographic column.
- Assessing monolayer permeability using elution profiles of optical tracers (blue dextran, sodium fluorescein, cyanocobalamin).
- Microfilament structure analysis using rhodamine-phalloidin staining; testing the effect of ketoconazole, a cytochrome P-450 inhibitor.
Main Results:
- Perilla ketone (PK) perfusion significantly increased endothelial permeability to sodium fluorescein and cyanocobalamin by over 50% within 15 minutes.
- PK-induced permeability changes were rapid, reversible upon PK removal, and disrupted endothelial actin microfilaments.
- Ketoconazole did not block PK's effect, suggesting cytochrome P-450 is not required for PK to increase vascular permeability.
Conclusions:
- Perilla ketone (PK) directly and rapidly increases endothelial cell permeability in vitro.
- PK disrupts endothelial actin cytoskeleton, contributing to its toxic effects.
- PK toxicity does not appear to be mediated by cytochrome P-450 activation.