Bronchial Contraction Research Articles

Different Effects of the K+ Channel Blockers 4‐Aminopyridine and Charybdotoxin on Sensory Nerves in Guinea‐pig Lung

In the isolated guinea-pig bronchus, the potassium channel blocking agent 4-aminopyridine (10(-4) M) caused a contraction which was abolished by capsaicin tachyphylaxis, suggesting involvement of sensory neuropeptides. Charybdotoxin (10(-8), 5 x 10(-8) M), which is a potent blocker of the high-conductance Ca(2+)-activated K+ channel in smooth muscle, caused slowly developing and long lasting bronchoconstriction, which was resistant to capsaicin tachyphylaxis. Neither 4-aminopyridine (10(-3), 10(-4) M) nor charybdotoxin (10(-8), 5 x 10(-8) M) had any significant effect on the bronchoconstriction induced by electrical field stimulation. Furthermore, charybdotoxin had no significant influence on the inhibitory effect of the alpha 2-adrenoceptor agonist SKF 35886 (5 x 10(-7) M) on the bronchoconstriction induced by electrical field stimulation. In the isolated perfused guinea-pig lung, 4-aminopyridine (3 x 10(-5) -10(-3) M) caused bronchoconstriction and enhanced both basal and (at 3 x 10(-5) M) vagal nerve stimulation-evoked calcitonin gene-related peptide outflow from pulmonary sensory nerves. In conclusion, 4-aminopyridine stimulated capsaicin-sensitive sensory neurons and enhanced the sensory activation induced by vagal nerve stimulation in guinea-pig lung. Charybdotoxin, on the other hand, caused bronchial contraction independently of capsaicin-sensitive nerves.

Low concentration endothelin-1 enhanced histamine-mediated bronchial contractions of guinea pigs in vivo

Endothelin-1 (ET-1) represents one of the most potent constrictors of guinea pig bronchial smooth muscle. In our study, various doses of aerosolized ET-1 increased airway resistance. But a very low concentration of ET-1 (10 −12M) could not change airway resistance. We examined low doses of ET-1 enhanced the sensitivity of bronchial contractions to histamine. ET-1 may play one of the most important roles in contributing directly or indirectly to the pathogenesis of bronchial asthma.

In vitro bronchial hyperresponsiveness after lung transplantation in the rat.

In this study, we examined the effect of unilateral lung transplantation on in vitro responses of bronchial smooth muscle to electrical field stimulation (EFS) and exogenous acetylcholine (ACh). In syngeneic Lewis rats, we compared the contractile responses of the left hilar transplanted bronchus with that of the nontransplanted right bronchus and the left and right bronchi from control and sham-operated animals. Atropine-sensitive bronchial contraction to EFS was greater on left transplanted than on right nontransplanted bronchi at all frequencies and voltages used. In control and sham-operated animals, contractile responses to EFS were identical on left and right sides and similar to the responses of the right nontransplanted bronchus of the lung transplant recipient. Concentration-response curves to exogenous ACh were similar on transplanted and all control bronchi. Intramural cholinergic nerve cell bodies reactive for acetylcholinesterase were present in the transplanted bronchus. We conclude that EFS-induced cholinergic contraction of rat bronchial smooth muscle is enhanced after lung transplantation. Enhancement is not a result of medication, inflammation, or rejection, absent from our syngeneic transplant, or of upregulation of muscarinic post-junctional receptors because the response to exogenous ACh was normal. Hence, an increased release of ACh from cholinergic nerve terminals may occur, due either to hyperexcitability of postganglionic parasympathetic nerves or to loss of inhibitory mechanism resulting from interruption of neural connections with the central nervous system.

Selective inhibition by dactinomycin of NANC sensory bronchoconstriction and [125I]NKA binding due to NK-2 receptor antagonism.

In the present study, dactinomycin (10(-5) M) inhibited the non-adrenergic, non-cholinergic bronchoconstriction upon antidromic vagal nerve stimulation (1 Hz for 1 min) in the isolated perfused guinea-pig lung by 84%. The release of calcitonin gene-related peptide was unchanged, however, suggesting a postjunctional action. Dactinomycin (10(-5), 5 x 10(-5) M) also reduced non-adrenergic non-cholinergic bronchial contractions (maximally by 75%) induced by electrical field stimulation or capsaicin, while the cholinergic component and non-adrenergic non-cholinergic relaxation remained intact. The neurokinin-2 receptor antagonist L-659,877 (10(-6) M) had a similar effect as dactinomycin, inhibiting the non-adrenergic non-cholinergic bronchial contractions by 69%, while the neurokinin-1 receptor antagonist CP-96,345 (10(-6) M) had no effect. The bronchoconstriction evoked by neurokinin A, the selective neurokinin-2 receptor agonist Nle10neurokinin A (4-10) and capsaicin was markedly inhibited by dactinomycin while the contraction induced by substance P (SP), the selective neurokinin-1 receptor agonist Sar9Met(O2)11SP, endothelin-1 and acetylcholine was not affected. In autoradiographic experiments on guinea-pig lung, [125I]neurokinin A-labelled sections showed dense binding in the bronchial smooth muscle layer. Dactinomycin inhibited the specific binding of [125I]neurokinin A in a concentration-dependent manner (IC50 = 6.3 x 10(-6) M) and 66% of [125I]neurokinin A total binding was inhibited by 10(-4) M dactinomycin. In the rat colon, [125I]neurokinin A binding to neurokinin-2 sites on circular smooth muscle was inhibited by dactinomycin with an IC50 value of 7.9 x 10(-6) M. Dactinomycin failed to reduce increased nerve-evoked contractions or those caused by Nle10neurokinin A (4-10) per se in the rat vas deferens, which are considered to be mediated by neurokinin-2 receptor activation. In the rat portal vein, dactinomycin did not influence the contractions caused by the neurokinin-3 selective agonist Pro7neurokinin B. In conclusion, dactinomycin selectively inhibited neurokinin-2 receptor activation in guinea-pig lung and rat colon, but not in rat vas deferens, which may depend on the existence of different neurokinin-2 receptor subtypes. Neurokinin A is most likely the main endogenous excitatory non-adrenergic non-cholinergic transmitter in guinea-pig bronchi.

Acute severe asthma: Part 1. Pathophysiology and clinical assessment.

Asthma manifests itself in a two-phase reaction. An early phase, causing bronchial muscle contraction, occurs within minutes after an antigenic stimulus. Within 6 to 18 hours, a late asthmatic response begins. This response is associated with cellular infiltration of the bronchial submucosa by eosinophils, neutrophils, and lymphocytes. The late asthmatic response is most effectively reversed by the use of corticosteroids. Predicting patient outcome solely on the basis of either physical findings, or pulmonary function tests, or historical events is frequently inadequate. Historical events, clinical assessments, and laboratory and pulmonary function measurements are best used in a complementary and integrated fashion to determine appropriate disposition of the acutely asthmatic patient.

Inhibition of Neutral Endopeptidase Potentiates Bronchial Contraction Induced by Immune Response in Guinea PigsIn Vitro

To study the role of tachykinins and neutral endopeptidase (NEP), an enzyme that degrades tachykinins, in the immune response in the airways of guinea pigs sensitized to ovalbumin (OVA), we examined the bronchial contractile response to OVA by inhibiting NEP in vitro. After incubating bronchial tissues with the NEP inhibitors phosphoramidon and thiorphan, we added 10(-5)% (10 micrograms/ml) OVA. Phosphoramidon and thiorphan (10(-5) M) significantly maintained the contraction that followed the peak contraction. In the next stages of the experiment, when the contraction induced by 10(-5)% OVA reached a plateau and began to relax, we added 10(-5) M phosphoramidon. Phosphoramidon inhibited the relaxation and significantly potentiated the contraction. In tissues treated with 10(-5) M capsaicin to deplete tachykinins, phosphoramidon did not potentiate the OVA-induced contraction, but substance P (10(-6) M) caused contraction. These results suggest that the immune response causes the release of tachykinin-like substances from capsaicin-sensitive nerves to induce bronchial contraction in part. To confirm the mediators that cause the release of the tachykinin-like substances from the bronchus, we also examined whether phosphoramidon potentiates the effect of leukotriene C4 (LTC4), serotonin, histamine, and platelet-activating factor on bronchial contraction. When the contractions induced by these agonists reached a plateau and began to relax, we added phosphoramidon. Phosphoramidon inhibited the relaxation and significantly potentiated the contractile response to 10(-5) M LTC4, and it significantly reduced the relaxing rate of the 10(-6) M serotonin-induced contraction. However, it did not change the effect of histamine and platelet-activating factor.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of preganglionic nerve evoked cholinergic contractions of the guinea pig bronchus

We compared cholinergic bronchial muscle contractions induced by vagus nerve (preganglionic) stimulation (VNS) with those induced by electrical field (postganglionic) stimulation (EFS). When normalized to their respective maximum response, the frequency-response curves (10 s trains) between 4 and 16 Hz were similar between VNS and EFS; however, at frequencies of 0.1–2 Hz, and at frequencies greater than 32 Hz, the VNS contractions were significantly less than EFS. When contractions elicited by 100 pulses were examined, it was found that the responses to VNS were maximal at 10–30 Hz then declined significantly to 82-35% of maximal between 40 and 200 Hz, whereas the response to EFS was essentially unchanged at frequencies up to 60 Hz and declined only to 72% of maximal up to 200 Hz. At frequencies as low as 20 Hz, the contractions evoked by VNS faded to 45 ± 9% of the peak contraction during 60 sec of continuous stimulation, whereas those evoked by 60 sec continuous EFS remained constant. This fade observed during prolonged VNS was not blocked by the antagonists, pirenzepine and AFDX-116, at concentrations selective for M1 and M2 muscarinic receptors, respectively; nor was the fade blocked by pre-treatment with indomethacin, propranolol, phentolamine, or choline. At frequencies greater than 10 Hz, the amplitude of the preganglionic compound action potential also faded during repetitive stimulation. The results support the hypothesis that the airway ganglion neurons innervating guinea pig bronchial smooth muscle effectively filter preganglionic stimuli, especially at low and relatively high frequencies. During continuous vagus nerve stimulation, preganglionic mechanisms may also play a role in limiting the ultimate output of airway ganglia.

The effect of cetirizine on the human isolated bronchus: Interaction with salbutamol

The effects of cetirizine (C), a new generation, nonsedative, H 1-antihistamine drug, were studied on human isolated bronchi. C is a potent antagonist of the bronchial muscle contraction induced by histamine, irrespective of whether C is administered by cumulative addition or prophylactically. In the former case, this effect of C was significant at a concentration of 3 × 10 −8 mol/L and was maximal at a concentration of 10 −5 mol/L. The −log of concentration of C causing 50% of the maximal effect induced was 7.30 ± 0.05 (n = 6), and the effect produced was not significantly modified by bronchial epithelium removal. When C was administered prophylactically, the concentration-response curves to histamine were displaced to the right, but the reduction of maximum histamine response suggests a noncompetitive type of antagonism. C is devoid of notable anticholinergic effects. At concentrations of 10 −8 to 10 −7 mol/L, C proved capable of enhancing the relaxant effect produced by salbutamol (10 −7 to 3 × 10 −7 mol/L) on human isolated bronchi that had been contracted by either histamine or acetylcholine (ACh). The synergy appeared to be additive or potentiating, depending on salbutamol (SAL) concentrations. Under similar conditions, mepyramine does not potentiate the effects of SAL against histamine. Finally, at concentrations of 10 −8 to 10 −6 mol/L, C reduced the functional antagonism observed between SAL and ACh, as can be observed by the increase, in the presence of C, of the maximal relaxant effect of SAL on contractions produced by 10 −3 mol/L of ACh. We may, therefore, conclude that C appears to be a specific antihistamine on human isolated bronchi and that it appears to potentiate the bronchodilator effect of SAL on this preparation.

Antigen-Induced Contraction of Human Isolated Lung Preparations Passively Sensitized with Monoclonal IgE: Effects of Indomethacin

Human isolated lung preparations were passively sensitized using mouse monoclonal dinitrophenyl (DNP)-specific IgE antibodies. The contractile response to antigen (DNP-bovine serum albumin; DNP-BSA) was approximately 80% of the histamine response (50 microM: 0.20 +/- 0.03 g/mm2) in bronchial muscle preparations. In passively sensitized pulmonary vascular and parenchymal preparations, the contraction to antigen was negligible. Indomethacin (1.7 microM; 30 min) did not alter the contractile response to DNP-BSA (5 micrograms/ml) in bronchial tissues. In passively sensitized bronchial muscle preparations stimulated with DNP-BSA, there was a significant increase (2-fold) in prostanoid production. This production was inhibited by indomethacin. These data suggest that endogenous prostaglandins may not play a role in the regulation of human isolated bronchial muscle contraction to antigen in vitro.

13 Inhibition of human bronchial contraction to 15-HETE by a leukotriene synthesis inhibitor and LTD 4 antagonist

13 Inhibition of human bronchial contraction to 15-HETE by a leukotriene synthesis inhibitor and LTD 4 antagonist

Corticosteroids in asthma

The treatment of asthma aims to keep the airways as normal as possible. They become narrowed because of mucosal inflammation and contraction of bronchial muscle. This mucosal inflammation is present even in the absence of symptoms,1 and probably causes the bronchial hyperreactivity so characteristic of asthma.2 If it is not treated adequately, the narrowing of the airways may become permanent.3 Bronchodilators treat symptoms but have no effect on the underlying inflammation. The early use of anti-inflammatory drugs might reverse this sequence, although there is no direct evidence that it alters the natural history of the disease.

CI-959, a New, Potential Antiallergic Drug, Inhibits Mediator Release from Lung and Contractions of Human Airways in vitro

Many symptoms of the immediate allergic response can be attributed to the synthesis/release and subsequent actions of histamine and metabolic products of arachidonic acid oxidation. CI-959 [5-methoxy-3-(1-methylethoxy)-N-1H-tetrazole-5-yl-benzo(b) thiophene-2-carboxamide], a new, potential antiallergic drug, inhibited the release of histamine, immunoreactive sulfidopeptide leukotrienes C4, D4 and E4 and immunoreactive thromboxane B2 from immunologically activated guinea-pig and human lung cells in vitro. The IC50s of CI-959 using guinea-pig lung were: histamine, 0.8 +/- 1.4 microM; leukotriene, 0.7 +/- 1.6 microM and thromboxane, 9.6 +/- 3.3 microM. Using human lung the IC50s were: 2.3 +/- 1.3 microM for histamine; 0.3 +/- 5.1 microM for leukotriene, and 0.3 +/- 2.6 microM for thromboxane. CI-959 caused a concentration-dependent inhibition of anti-IgE-induced contractions of human bronchial muscle. Mean percent inhibitions were 45, 65 and 96 at 1, 3 and 10 microM, respectively. Cromolyn, 10 microM, inhibited bronchial contractions only 42%. The ability of CI-959 to inhibit these immunologically induced contractions indicates that the release of all mediators responsible for bronchoconstriction was effectively inhibited. These data suggest that CI-959 may be effective in preventing the development of symptoms directly related to inflammatory mediator release in a variety of allergic and inflammatory states.

気管支ぜん息児における連続性ラ音の音響学的解析

Continuous rales heard during spontaneous attacks of bronchial asthma (natural attacks) and those of exercise-induced asthma (EIA) in children were analyzed sonographically using a sound spectrograph. We analyzed rales simultaneously recorded at three different locations, i. e. on the left and right sides of the chest and on the neck. Natural attacks were found in 68 patients and EIA in 31.The number and the duration of rales per respiratory cycle significantly increased in patients with natural attacks and in those with EIA according to the severity of the attack and the degree of decrease of FEV1. 0. In both natural attacks and EIA, the number of sites of rales were found to increase in a correlation with the degree of attacks and a decrease in pulmonary function.When comparing rales of natural attacks and EIA the fundamental frequency of the rales was significantly higher in EIA. This suggests that the location of bronchial contraction in EIA is more central than in natural attacks.The highest prevelence of rales among the three different pick-up points was on the neck. About 90% of rales were recorded on the neck, and about 90% of rales recorded on the chest were also simultaneously detected on the neck. We conclude that the neck is a important location for auscultating rales and rales are strongly transmitted to the neck than over the other side of the chest.Rales of short duration occurring in children with natural attacks and EIA were different from the squawks that were classified as discontinuous rales in adults with interstitial pneumonitis. The duration, number of overtones and timing of onset were all different in those two groups. We conclude that the mechanism of production may be nearly equal to that of continuous rales.

Adenosine selectively inhibits noncholinergic transmission in guinea pig bronchi.

The neuromodulatory action of adenosine and ATP was investigated in isolated guinea pig bronchial strip chain preparations contracted with electrical field stimulation. The tissues were placed in organ baths containing physiological salt solution and stimulated at 8-Hz frequency, 0.5-ms pulse duration, and 30 V (approximately 100 mA) for 5 s. Electrical field stimulation evoked a biphasic contraction of bronchial muscle, consisting of an initial contraction followed by a sustained contraction, which was mediated by intramural cholinergic and noncholinergic nerve stimulations, respectively. Adenosine, at concentrations greater than M, caused a concentration-dependent inhibition in the height of the noncholinergically mediated contraction, accompanied by a very weak inhibition on the cholinergically mediated response. ATP (10(-5) to 3 x 10(-3) M) also produced a similar inhibitory effect on the noncholinergically mediated contraction, but the inhibitory potency was less than that of adenosine. The inhibitory response to adenosine was enhanced by the pretreatment with dipyridamole (2 x 10(-6) M) but antagonized with aminophylline (10(-5) M). Contractions of bronchial muscle evoked by exogenous acetylcholine (2 x 10(-6) M) or substance P (2 x 10(-7) M) were significantly inhibited by the adenosine (3 x 10(-4) M) pretreatment. These data suggest that in isolated guinea pig bronchi adenosine selectively inhibits noncholinergic neurotransmission through prejunctional P1-purinoceptors.

Inhibition of cholinergic and non-adrenergic, non-cholinergic bronchoconstriction in the guinea pig mediated by neuropeptide Y and α2-adrenoceptors and opiate receptors

The mechanisms underlying the regulatory influence of neuropeptide Y (NPY) and of α 2-adrenoceptor and opiate receptor activation on cholinergic and excitatory non-adrenergic, non-cholinergic (e-NANC) bronchial contractions studied in guinea pig hilus bronchi in vitro. NPY inhibited both the cholinergic and e-NANC bronchial contractions evoked by field stimulation. The NPY attenuation of the e-NANC contraction could not be antagonized by the α 2-antagonist, idazoxan, or naloxone. UK 14,304 a specific α 2-agonist, also reduced the two nervous components of bronchial contraction and this action was inhibited by idazoxan. NPY and UK 14,304 exerted a minor influence on the bronchial smooth muscle tone per se or on contractions evoked by acetylcholine or neurokinin A. This suggested that the inhibitory responses were caused by a prejunctional action reducing the release of transmitter substances from sensory and cholinergic nerve endings. Furthermore NPY (10 −7 M) seemed to be more potent to inhibit both contractile components than noradrenaline (10 −6 M) in the presence of propranolol (3 × 10 −6 M). Morphine was able to reduce the e-NANC response via a naloxone-sensitive mechanism. The capsaicin-evoked bronchoconstriction and the bronchodilator NANC effect evoked by field stimulation were, however, not influenced by UK 14,304. It is concluded that NPY, α 2-receptor and opiate receptor activation inhibit the release of sensory transmitters evoked by field stimulation but not by capsaicin.

A muscarinic receptor subtype modulates vagally stimulated bronchial contraction.

An in vitro preparation was developed to study vagus nerve-stimulated (preganglionic) and field-stimulated (post-ganglionic) contraction of the rabbit main stem bronchus and to compare the inhibitory effects of muscarinic antagonists on that contraction. The maximal contractile responses (20 V, 0.5 ms, 64 Hz) for either field or vagal stimulation were completely abolished by atropine (60 nM). Hexamethonium (0.1 mM) abolished the response to vagal stimulation but did not affect the field-stimulated response. To compare the effectiveness of atropine and pirenzepine as antagonists at the nerve-smooth muscle junction, inhibition studies of field-stimulated contractions were performed. Pirenzepine was 102- to 178-fold less potent than atropine when compared at the inhibitory concentration of antagonist that produced 25, 50, and 75% inhibition (IC25, IC50, and IC75, respectively), indicating that the muscarinic receptor at the nerve-smooth muscle junction is a muscarinic receptor with low affinity for pirenzepine (M2 subtype). Atropine had similar inhibitory effects on vagal- and field-stimulated contractions. In contrast, pirenzepine was more potent in inhibiting vagally stimulated contraction than field-stimulated contraction, especially at the IC25 where pirenzepine was only 8- to 22-fold less potent than atropine in inhibiting vagally stimulated contraction. These data suggest that an M1 subtype of muscarinic receptor modulates excitatory neurotransmission through bronchial parasympathetic ganglia.

Evidence for two independent modes of activation of the ‘efferent’ function of capsaicin-sensitive nerves

Field stimulation (10 Hz for 10 s, 0.5 ms pulse width, 60 V) of the guinea-pig isolated main bronchi (atropine plus indomethacin in the bath) produced reproducible contractions which were abolished by tetrodotoxin or in vitro capsaicin desensitization. These responses were almost abolished by ω-conotoxin GVIA (CTX), a peptide modulator of neuronal calcium channels which, however, did not affect the bronchial contraction due to neurokinin A or to capsaicin. Field stimulation (10 Hz for 2.5 s, 1 ms, 60 V) of the electrically driven, isolated guinea-pig left atria excised from reserpine-pretreated animals (atropine in the bath) produced a delayed positive inotropic response which was abolished by tetrodotoxin or in vitro capsaicin desensitization. This response was abolished by CTX, which did not affect the response to exogenous calcitonin gene-related peptide nor that to capsaicin. These findings indicate that CTX-sensitive mechanisms (presumably Ca channels regulating the release of transmitters) are activated upon antidromic invasion of sensory terminals and consequent production of the ‘efferent’ response while the activation of sensory nerve endings by capsaicin occurs through CTX-resistant mechanisms.

The toxic effect of an AChE-inhibitor on the cholinergic nervous system in airway smooth muscle

Excessive cholinergic stimulation of presynaptic muscarinic cholinergic receptors, due to complete inhibition of acetylcholinesterase (AChE) by O-(1,2,2-trimethylpropyl)-methyl-phosphonofluoridate (soman), reduced the release of acetylcholine (ACh) from cholinergic nerves in rat bronchi by almost 25%. Furthermore, long-term (40 h) exposure by inhalation of soman (0.45–0.63 mg/m 3) reduced the contraction of bronchi induced by ACh by approximately 70%. This is probably due to reduction of the number of muscarinic cholinergic receptors, since there was a reduction in the binding capacity ( B max) of [ 3H]QNB by 40%, without any changes in the dissociation constant ( K d).

Regional differences in the reactivity of guinea-pig airways

Regional differences in the sensitivity of guinea-pig airways to cholinergic and beta-adrenoceptor stimulation as well as Schultz-Dale responses were studied in vitro. Carbachol and isoproterenol produced a 27% and 39% greater response, respectively, in the trachea as compared to the bronchus. Contractions to histamine in tracheal and bronchial tissues were significantly less than those evoked by carbachol (P less than 0.05); however, lung parenchymal strips did not show any difference in maximal contractility to these agonists. Responses to antigen in airways from sensitized guinea-pigs or responses to anaphylactic mediators, LTD4 and PGD2, in normal airways did not show any differences in maximum tissue contractility (trachea and bronchus). Among the airway tissues examined, lung parenchymal strips were more responsive to these mediators as compared to tracheal and bronchial tissues. Threshold and sub-threshold concentrations of LTD4 had no effect on tracheal and bronchial airway contractions to PGD2; however, PGD2-induced contractions of the lung parenchymal strip were significantly inhibited in the presence of LTD4 (P less than 0.0001). Our data suggest that: (1) considerable regional differences exist in the reactivity of guinea-pig airways; (2) the maximum responses of bronchial airways to cholinergic and beta-adrenergic stimuli appear less than that observed in the trachea; (3) small peripheral airways appear more responsive to some mediators of anaphylaxis than are the larger central airways. Therefore, it would appear that the trachea and bronchus are more responsive to cholinergic stimuli, while the lung parenchyma is more responsive to PGD2, LTD4 and also to in vitro anaphylaxis.(ABSTRACT TRUNCATED AT 250 WORDS)

Peripheral effects of opioid drugs on capsaicin-sensitive neurones of the guinea-pig bronchus and rabbit ear.

The effect of a potent opioid agonist, [D-Met2, Pro5]-enkephalinamide was investigated on two responses involving capsaicin-sensitive afferent neurones, namely, atropine-resistant contractions of the guinea-pig bronchus evoked by electrical field stimulation and the nociceptor stimulation to intraarterial injections of acetylcholine or capsaicin into the vascularly isolated rabbit ear. The hypotheses to be tested were whether (a) opioid receptor activation may inhibit mediator release from primary afferent neurones and (b) the opioid could exert an analgesic effect at a peripheral site of action. Non-cholinergic contractions of the guinea-pig isolated main bronchi due to electrical stimulation were concentration-dependently inhibited by [D-Met2, Pro5]-enkephalinamide (10 nM-1 microM). This effect was abolished by naloxone (1 microM). Naloxone alone induced no change in the stimulation-evoked contractions of the bronchus, indicating that no endogenous opioid control was present. Substance P and neurokinin A induced bronchial contractions that were not influenced by [D-Met2, Pro5]-enkephalinamide. This indicates that [D-Met2, Pro5]-enkephalinamide inhibits electrically-evoked bronchial contractions by reduced mediator release from capsaicin-sensitive sensory nerve endings, since these contractions are most probably brought about by tachykinins, released from afferent neurones. Capsaicin-induced bronchial contractions were in contrast to electrical stimulation not influenced by [D-Met2, Pro5]-enkephalinamide which suggests a different site of action. The activation of sensory neurones in the rabbit ear by i.a. injection of acetylcholine and capsaicin was not reduced under infusion of [D-Met2, Pro5]-enkephalinamide (1 and 10 microM) or lofentanil (1 and 10 microM).(ABSTRACT TRUNCATED AT 250 WORDS)