title: Group B Streptococcus and E. coli LPS-induced NO-dependent hyporesponsiveness to noradrenaline in isolated intrapulmonary arteries of neonatal piglets.
abstract: 1. The effects of endotoxin (E. coli lipopolysaccharide, LPS) and heat inactivated group B Streptococcus (GBS) were studied on the contractile responses to noradrenaline (NA) in isolated pulmonary arteries and on the activity of the constitutive and inducible nitric oxide synthase (NOS) in lung fragments of neonatal piglets. 2. Short-term (< or = 5 h) incubation with LPS (1 micrograms ml-1) or GBS (3 x 10(7) colonies forming units ml-1) did not modify the vascular responsiveness to NA (10(-8) M-10(-4) M) in isolated intrapulmonary arteries. However, long-term incubation (20 h) with LPS or GBS produced a significant reduction in the maximal contractile responses and shifted the concentration-response curve for NA downwards. 3. Endothelium removal or the cyclo-oxygenase inhibitor meclofenamate (10(-5) M) did not affect the GBS- and LPS-induced hyporesponsiveness to NA. 4. The presence of the nitric oxide (NO) precursor, L-arginine (10(-5) M), 30 min prior to the contractility challenge increased the LPS- and GBS-induced pulmonary vascular hyporesponsiveness to NA. In contrast, the addition, prior to the challenge with NA, of the NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME, 10(-4) M) or coincubation with dexamethasone (3 x 10(-6) M), a potent inhibitor of the induction of NOS, or with the protein synthesis inhibitor cycloheximide (10(-5) M) completely restored the reactivity to NA in LPS- and GBS-treated pulmonary arteries. 5. The incubation for 20 h of lung fragments with LPS and GBS produced a significant increase in the Ca2+-independent (inducible) NOS activity determined by the conversion of radiolabelled L-arginine to citrulline, but did not modify the constitutive NOS activity. This NOS induction was abolished by coincubation with dexamethasone (3 X 10-6 M).6. These results demonstrated that prolonged incubation with GBS and LPS causes an induction of NOS activity which results in a reduced vascular responsiveness to NA in pulmonary arteries of neonatal piglets. Thus, induction of NOS seems to be responsible for the delayed pulmonary vascular hyporesponsiveness induced by GBS (a Gram-positive) and E. coli (a Gram-negative), the most common causal agents of neonatal sepsis.
authors.affiliation: Department of Pharmacology, School of Medicine, Universidad Complutense, Madrid, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Villamor
authors.forename: F
authors.initials: F
authors.lastname: Pérez-Vizcaíno
authors.forename: T
authors.initials: T
authors.lastname: Ruiz
authors.forename: J C
authors.initials: JC
authors.lastname: Leza
authors.forename: M
authors.initials: M
authors.lastname: Moro
authors.forename: J
authors.initials: J
authors.lastname: Tamargo
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004790:Enzyme Induction / Q000187:drug effects; D004926:Escherichia coli / Q000472:pathogenicity*; D008070:Lipopolysaccharides / Q000633:toxicity*; D008168:Lung / Q000098:blood supply / Q000187:drug effects / Q000201:enzymology; D008297:Male; D008469:Meclofenamic Acid / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D019331:NG-Nitroarginine Methyl Ester; D009569:Nitric Oxide / Q000037:antagonists & inhibitors / Q000378:metabolism; D019001:Nitric Oxide Synthase; D009638:Norepinephrine / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000187:drug effects; D051381:Rats; D013292:Streptococcus agalactiae / Q000472:pathogenicity*; D013552:Swine; D014655:Vascular Resistance / Q000187:drug effects
chemical_list: D008070:Lipopolysaccharides; D009569:Nitric Oxide; D008469:Meclofenamic Acid; D003907:Dexamethasone; D001120:Arginine; D003513:Cycloheximide; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D019331:NG-Nitroarginine Methyl Ester; D009638:Norepinephrine