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Continuous measurements of plasma protein extravasation with microdialysis after various inflammatory challenges in
V V Iversen1, A Bronstad, E-A B Gjerde
1Department of Physiology, Jonas Lies vei 91, N-5009 Bergen, Norway. vegard.iversen@fys.uib.no
Abstract:
This study describes the use of microdialysis technique for continuous measurement of plasma protein extravasation (PPE) in rat and mouse skin with drug application either intravenously or via the microdialysis fiber. Hollow plasmapheresis fibers (3-cm length, 0.4-mm diameter, cutoff 3,000 kDa) were placed subcutaneously on the back of anesthetized mice and rats. Intravenous injection of dextran (Macrodex, 60 mg/ml) increased PPE by 355% from baseline within 30 min in rats with ligated kidneys (n = 6; P < 0.05) but not in animals with intact kidneys. Phalloidin (500 microg/kg iv 40 min before dextran, n = 6; P < 0.05) did not change the response to dextran in either group. Animals receiving PGE1, compound 48/80 (mice), paclitaxel, docetaxel, and cremophor EL via the microdialysis fiber were also provided with a control fiber receiving vehicle. Both rats and mice had constant PPE in the control fiber, and there was no change in PPE in the NaCl-treated groups (rats, n = 4; mice, n = 6). Application via the fiber of PGE1 (20 microg/ml), compound 48/80 (mice; 4 mg/ml), and docetaxel (0.5 mg/ml) increased PPE compared with baseline within 60 min by 139% (n = 6; P < 0.05), 273% (n = 6; P < 0.05), and 325% (n = 5; P < 0.05), respectively. Phalloidin alone did not increase PPE (n = 5; P < 0.05). Pretreatment with phalloidin did not inhibit the increase after PGE1 or compound 48/80 but inhibited that after docetaxel (n = 6). Paclitaxel (0.6 mg/ml, n = 5) or vehicle (Cremophor) (n = 5) gave no increase in PPE. The results demonstrate that microdialysis can be used to continuously measure changes in PPE after inflammatory challenges in skin of rats and mice.
Insights
This study demonstrates microdialysis for continuous measurement of plasma protein extravasation (PPE) in rodent skin. The technique effectively quantifies drug-induced changes in PPE, aiding inflammatory research.
Area of Science:
- Pharmacology
- Physiology
- Biomedical Engineering
Background:
- Plasma protein extravasation (PPE) is a key indicator of inflammation and vascular permeability.
- Accurate and continuous measurement of PPE is crucial for understanding inflammatory processes and drug effects.
- Existing methods may lack the resolution or continuous monitoring capabilities needed for dynamic studies.
Purpose of the Study:
- To establish and validate the microdialysis technique for continuous measurement of plasma protein extravasation (PPE) in rat and mouse skin.
- To assess the impact of various drug applications (intravenous and local) on PPE.
- To evaluate the efficacy of microdialysis in response to inflammatory stimuli and drug challenges.
Main Methods:
- Utilized hollow plasmapheresis fibers (3 cm length, 0.4 mm diameter, 3,000 kDa cutoff) implanted subcutaneously in anesthetized rats and mice.
- Administered substances intravenously (dextran, phalloidin) or locally via the microdialysis fiber (PGE1, compound 48/80, paclitaxel, docetaxel, cremophor EL).
- Measured continuous changes in PPE in response to drug application and inflammatory challenges, comparing with control groups.
Main Results:
- Intravenous dextran significantly increased PPE in rats with ligated kidneys, but not in those with intact kidneys.
- Local application of PGE1, compound 48/80, and docetaxel via microdialysis fibers significantly increased PPE in both rats and mice.
- Phalloidin pretreatment inhibited the docetaxel-induced increase in PPE, while paclitaxel and cremophor EL did not affect PPE.
Conclusions:
- Microdialysis is a reliable method for continuous, real-time measurement of plasma protein extravasation in rodent skin.
- The technique can effectively detect and quantify PPE changes induced by various inflammatory agents and drugs.
- This method provides a valuable tool for pharmacological research, particularly in studying vascular permeability and drug efficacy in inflammatory conditions.
