Temporal variation in endotoxin-induced vascular hyporeactivity in a rat mesenteric artery organ culture model.
Endotoxin-induced vascular hyporeactivity to phenylephrine (PE) is well described in rodent aorta, but has not been investigated in smaller vessels in vitro. Segments of rat superior mesenteric artery were incubated in culture medium with or without foetal bovine serum (10%) for 6, 20 or 46 h in the presence or absence of bacterial lipopolysaccharide (LPS; 1 - 100 microg ml(-1)). Contractions to PE were measured with or without nitric oxide synthase (NOS) inhibitors: L-NAME (300 microM), aminoguanidine (AMG; 400 microM) 1400W (10 microM) and GW273629 (10 microM); the guanylyl cyclase inhibitor, ODQ (3 microM); the COX-2 inhibitor, NS-398 (10 microM). Contractile responses to the thromboxane A2 mimetic, U46619 were also assessed. In the presence of serum, LPS induced hyporeactivity at all time points. In its absence, hyporeactivity only occurred at 6 and 20 h. L-NAME and AMG fully reversed hyporeactivity at 6 h, whereas they were only partially effective at 20 h and not at all at 46 h. In contrast partial reversal of peak contraction was observed with 1400W (62% at 46 h), GW273629 (57% at 46 h) and ODQ (75% at 46 h). COX-2 inhibition produced no reversal. In contrast to PE, contractions to U46619 were substantially less affected by LPS. We describe a well-characterized reproducible model of LPS-induced hyporeactivity, which is largely mediated by the NO-cyclic GMP-dependent pathway. Importantly, long-term (2-day) production of NO via iNOS is demonstrated. Moreover, conventional doses of L-NAME and AMG became increasingly ineffective over time. Thus, the choice of inhibitor merits careful consideration in long-term models.
O'Brien
A J
AJ
The Centre for Clinical Pharmacology, Department of Medicine, University College London, Rayne Institute, University Street, London WC1E 6JJ, UK. alastair.o'brien@ucl.ac.uk
Wilson
A J
AJ
Sibbald
R
R
Singer
M
M
Clapp
L H
LH
D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / Q000494:pharmacology; D000818:Animals; D051546:Cyclooxygenase 2; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology; D006157:Guanosine Monophosphate / Q000378:metabolism; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors; D007527:Isoenzymes / Q000037:antagonists & inhibitors; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D008638:Mesenteric Arteries / Q000187:drug effects* / Q000502:physiology*; D023421:Models, Animal; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D009578:Nitrobenzenes / Q000494:pharmacology; D009924:Organ Culture Techniques; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D010656:Phenylephrine / Q000494:pharmacology*; D011451:Prostaglandin-Endoperoxide Synthases; D051381:Rats; D017207:Rats, Sprague-Dawley; D013449:Sulfonamides / Q000494:pharmacology; D013997:Time Factors; D014661:Vasoconstriction / Q000187:drug effects*; D014662:Vasoconstrictor Agents / Q000494:pharmacology*
D007527:Isoenzymes; D008070:Lipopolysaccharides; D020030:Nitric Oxide Donors; D009578:Nitrobenzenes; C110881:S-nitro-N-acetylpenicillamine; D013449:Sulfonamides; D014662:Vasoconstrictor Agents; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; D010656:Phenylephrine; D009569:Nitric Oxide; D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; D006157:Guanosine Monophosphate; D019001:Nitric Oxide Synthase; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D006162:Guanylate Cyclase; D010396:Penicillamine