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Hyperresponsiveness of airway smooth muscle to allergens and environmental factors has long been associated with the pathophysiology of asthma. Tryptase, a serine protease of lung mast cells, has been implicated as one of the mediators involved in the induction of hyperresponsiveness. As a consequence, tryptase inhibitors have become the subject of study as potential novel therapeutic agents for asthma. Secretory leukocyte protease inhibitor (SLPI) is a naturally occurring protein of human airways which exhibits anti-tryptase activity. To assess the potential therapeutic utility of SLPI in asthma, its effects were evaluated using in vitro and ex vivo models of airway hyperresponsiveness and compared with the effects of the small molecule tryptase inhibitor APC-366. Our results demonstrate that SLPI inhibits tryptase-mediated hyperresponsiveness in vitro and attenuates the hyperresponsiveness observed in airway smooth muscle from antigen-sensitized animals subjected to antigen exposure. The small molecule tryptase inhibitor APC-366 has a similar inhibitory effect. Thus, tryptase appears to be a significant contributor to the development of hyperresponsiveness in these models. To the extent that tryptase contributes to the development and progression of asthma, SLPI may posses therapeutic potential in this disease setting.

Neuropeptide Y (NPY) potentiates the contractile response of the rat caudal artery to adrenergic nerve stimulation in-vitro . The NPY Y 1 selective antagonist BIBP3226 ((R)-N 2 -(diphenylacetyl)-N-[(4-hydroxyphenyl)methyl]-argininamide), inhibited the vascular effects of NPY in rat caudal artery preparations in-vitro (IC 50 = 126 nM). BIBP3226 also inhibited the effects of the selective Y 1 agonist [Leu 31 ,Pro 34 ]NPY and completely abolished the effects of avian pancreatic polypeptide that was shown to be capable of potentiating neurogenic vasoconstriction in this preparation. These effects were reversible and are most likely mediated by the Y 1 receptor subtype since we failed to observe any functional evidence of a Y 2 receptor subtype in rat caudal artery. The caudal artery provides a useful functional assay for pharmacological analysis of NPY and NPY antagonists.

Pancreatic polypeptide (PP) is a member of the PP fold family of regulatory peptides. Studies have shown that neuropeptide Y, peptide YY, and PP increased gastrointestinal motility. The GI effects of neuropeptide Y and peptide YY were accompanied by an increase in mean arterial blood pressure; however, PP decreased mean arterial blood pressure. Cloning of a receptor of the neuropeptide Y family with high affinity for PP has been reported. This Y4 receptor is present in intestine, pancreas, and prostate, and its mRNA has been detected in brain and coronary artery. We found in vitro evidence of PP-mediated inhibition of arterial neurogenic vasoconstriction. We have also detected Y4 mRNA in rat peripheral arteries. These findings suggest a potential role for the Y4 receptor in regulating vascular tone.

The hormone melatonin potentiated contractile responses to adrenergic nerve stimulation in isolated ring segments of rat caudal artery. This effect was inhibited by the melatonin receptor antagonist luzindole but not by the serotonin 5-HT2 receptor antagonist ketanserin. Melatonin had no direct effects on vascular tone. Melatonin agonists potentiated contractile responses with a relative order of potency (2-iodomelatonin, EC50 = 0.6 nM; melatonin, EC50 = 4.7 nM; N-acetylserotonin, EC50 = 1.5 microM) that is consistent with the melatonin ML1 receptor subtype. Melatonin also potentiated contractions elicited by exogenous norepinephrine and produced its effects in the absence of an intact endothelium. These data suggest that melatonin acts on receptors in the smooth muscle. The caudal artery provides a useful functional assay for pharmacological analysis of melatonin receptors. Physiologically, melatonin may activate its receptors at night to influence thermoregulation in the rat by enhancing the effects of sympathetic input to the caudal artery.

Prejunctional effects of opioids were examined in the perfused mesentery of two species: the rat and rabbit. Use of agonists selective for subtypes of mu, delta, and kappa opioid receptors produced no effect on contractile responses to adrenergic nerve stimulation in the rat perfused mesentery, except for small effects of the kappa agonist EKC, which may be non specific. In contrast, mu, delta and kappa receptors appear to be present in the rabbit. The mu selective agonist, DAMGO, kappa agonist, ethylketocyclazocine, and delta agonists, DPDPE and [Leu5]-enkephalin, all produced significant inhibition of contractile responses to transmural nerve stimulation. The inhibitory effect was greatest for ethylketocyclazocine. To test the possibility that prejunctional activation of alpha 2 adrenoceptors with endogenous norepinephrine might decrease the activity of prejunctional opioid receptors in the rabbit, inhibitory effects of delta and kappa selective agonists were tested in the presence of 10(-7) M yohimbine. Inhibitory responses of the kappa selective agonist ethylketocyclazocine were enhanced, while that of delta selective agonists [Leu5]-enkephalin and DPDPE remained unchanged when yohimbine was present. Thus, the effects of opioids vary and depend on the tissue and receptor subtypes they act upon. Furthermore, the enhanced inhibitory effect of opioid receptor activation in the presence of yohimbine is not found for all opioid receptors.