Iloprost inhalation redistributes pulmonary perfusion and decreases arterial oxygenation in healthy volunteers.

BACKGROUND: Previous studies have shown that ventilation-perfusion matching is improved in the prone as compared with that in the supine position. Regional differences in the regulation of vascular tone may explain this. We have recently demonstrated higher production of nitric oxide in dorsal compared with ventral human lung tissue. The purpose of the present study was to investigate regional differences in actions by another vasoactive mediator, namely prostacyclin. The effects on gas exchange and regional pulmonary perfusion in different body positions were investigated at increased prostacyclin levels by inhalation of a synthetic prostacyclin analogue and decreased prostacyclin levels by unselective cyclooxygenase (COX) inhibition. METHODS: In 19 volunteers, regional pulmonary perfusion in the prone and supine position was assessed by single photon emission computed tomography using (99m)Tc macro-aggregated albumin before and after inhalation of iloprost, a stable prostacyclin analogue, or an intravenous infusion of a non-selective COX inhibitor, diclofenac. In addition, gas distribution was assessed in seven subjects using (99m)Tc-labelled ultra-fine carbon particles before and after iloprost inhalation in the supine position. RESULTS: Iloprost inhalation decreased arterial PaO(2) in both prone (from 14.2+/-0.5 to 11.7+/-1.7 kPa, P<0.01) and supine (from 13.7+/-1.4 to 10.9+/-2.1 kPa, P<0.01) positions. Iloprost inhalation redistributed lung perfusion from non-dependent to dependent lung regions in both prone and supine positions, while ventilation in the supine position was distributed in the opposite direction. No significant effects of non-selective COX inhibition were found in this study. CONCLUSIONS: Iloprost inhalation decreases arterial oxygenation and results in a more gravity-dependent pulmonary perfusion in both supine and prone positions in healthy humans.

Regional differences in nitric oxide-mediated vasorelaxation in porcine pulmonary arteries.

BACKGROUND: Several previous investigations have shown improved oxygenation when ventilator-treated patients with acute lung injury are turned prone. In a previous human study, we demonstrated higher Ca(2+)-dependent nitric oxide synthase (NOS) activity in dorsal than in ventral parts of the lung. The current investigation was designed to determine whether Ca(2+)-dependent NOS activity was different in dorsal and ventral porcine lung regions. In addition, possible differences in vascular responses to nitroprusside or secondary to acetylcholine- or bradykinin-stimulated NO production were studied in dorsal and ventral pulmonary arteries. METHODS: In the study, 20 pigs were used. Lung biopsies and pulmonary arterial rings were harvested from ventral and dorsal lung regions. NOS activity was determined by citrulline assay in the presence and absence of the calcium chelator ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid (EDTA) to discriminate between Ca(2+)-dependent and Ca(2+)-independent NOS activity. In organ baths, in submaximally contracted arterial rings, vasorelaxation induced by acetylcholine, bradykinin and nitroprusside was measured. RESULTS: Ca(2+)-dependent NOS activity was higher in dorsal parts (87.2 +/- 9.1 citrulline units) than in ventral parts (62.2 +/- 10.1 citrulline units, P < 0.05) of porcine lung. There was a greater relaxation in dorsal than in ventral pulmonary arterial rings induced by both acetylcholine and bradykinin. Nitroprusside relaxed both sites equally. CONCLUSIONS: Our results show that endothelial-derived NO is an important factor influencing the differences between dorsal and ventral lung regions in vasorelaxing activity in porcine pulmonary arteries. This finding provides an explanation for the improved oxygenation when patients with severe acute lung insufficiency are turned prone.

Interactions of volatile anesthetics with cholinergic, tachykinin, and leukotriene mechanisms in isolated Guinea pig bronchial smooth muscle.

UNLABELLED: We studied relaxation of airway smooth muscle by sevoflurane, desflurane, and halothane in isolated guinea pig bronchi. Ring preparations were mounted in tissue baths filled with physiological salt solution and continuously aerated with 5% CO(2) in oxygen. Electrical field stimulation induced contractions sensitive to tetrodotoxin, indicating nerve-mediated responses. These consisted of an atropine-sensitive cholinergic phase and a nonadrenergic noncholinergic (NANC) phase sensitive to SR48968, a neurokinin-2 receptor antagonist. Anesthetics were added to the gas aerating the tissue baths. Sevoflurane and desflurane at 1.0 minimum alveolar anesthetic concentration and halothane at 1.0-2.0 minimum alveolar anesthetic concentrations inhibited both cholinergic and NANC contractions to electrical field stimulation. None of the anesthetics affected responses to exogenously applied neurokinin A, a likely mediator of NANC contractions, suggesting prejunctional inhibition of NANC neurotransmission. The anesthetics did not affect the initiation of contractile responses to leukotriene C(4) (LTC(4)), a mediator of asthmatic bronchoconstriction. However, sevoflurane and desflurane both relaxed bronchi in a steady-state contraction achieved by LTC(4). Surprisingly, halothane did not relax LTC(4) contractions. Concerning LTC(4)-elicited bronchoconstriction, sevoflurane and desflurane were more potent airway smooth muscle relaxants in vitro. IMPLICATIONS: Halothane, sevoflurane, and desflurane attenuated airway smooth muscle tone via inhibition of cholinergic and nonadrenergic noncholinergic neurotransmission. Sevoflurane and desflurane reduced leukotriene C(4)-induced bronchoconstriction, whereas halothane did not. This indicates a beneficial role for sevoflurane and desflurane in asthmatics.

Relaxation by sevoflurane, desflurane and halothane in the isolated guinea-pig trachea via inhibition of cholinergic neurotransmission.

We have studied relaxation of airway smooth muscle by sevoflurane, desflurane and halothane in the isolated guinea-pig trachea. Ring preparations were mounted in tissue baths filled with physiological salt solution (PSS), aerated continuously with 5% carbon dioxide in oxygen. Electrical field stimulation (EFS) elicited cholinergic contractions that were abolished by tetrodotoxin, indicating nerve-mediated responses. Anaesthetics were added to the gas aerating the tissue baths. Halothane, sevoflurane and desflurane at 0.5-1.0 MAC markedly attenuated cholinergic contractions to EFS. Initiation of contractile responses to acetylcholine (ACh) were not affected by volatile anaesthetics, suggesting prejunctional inhibition (i.e. inhibition of acetylcholine release). When added to a maintained submaximal contraction to ACh, volatile anaesthetics induced relaxation, indicating postjunctional inhibition. We conclude that sevoflurane, desflurane and halothane inhibited postganglionic cholinergic neuroeffector transmission in the trachea. The effect was probably exerted via pre- and postjunctional mechanisms (i.e. inhibition of acetylcholine release and direct muscle actions). Sevoflurane and desflurane were more potent than halothane both pre- and postjunctionally.

Ethanol causes decrements in airway excretion of endogenous nitric oxide in humans.

Ethanol has previously been demonstrated to inhibit excretion of endogenous nitric oxide (NO) in exhaled air from experimental animals. The aim of the present study was to elucidate if this effect also occurs in human subjects. Healthy volunteers ingested ethanol (0.25 and 1 g kg-1, 20% in orange juice). Nitric oxide in exhaled air was determined by chemiluminescence. Single-breath analysis of exhaled air was performed and peak values of NO and end expiratory levels of NO and CO2 were determined. Ethanol induced dose-dependent decrements in exhaled nitric oxide. Thus, peak values for nitric oxide in exhaled air, in the first exhalation after breath-holding for 30 s, decreased to 56 +/- 10 and 37 +/- 12% of control 60 min after ingestion of ethanol at 0.25 and 1 g kg-1, respectively. Rinsing the oral cavity (including gargling) for 15 min with 20% ethanol in juice did not significantly influence NO in exhaled air. Heart rate blood pressure and end expiratory levels of CO2 were not significantly affected by ethanol ingestion. In conclusion, ethanol decreases levels of nitric oxide in exhaled air in humans, likely by inhibition of airway formation of nitric oxide. The results might be of importance in understanding effects of ethanol and other hydrocarbons.

Modulation of cholinergic and substance P-like neurotransmission by nitric oxide in the guinea-pig ileum.

1. The role of endogenous nitric oxide (NO) as a modulator of enteric neurotransmission was investigated in longitudinal muscle myenteric plexus (LMMP) preparations of guinea-pig isolated ileum. 2. In tissues previously incubated with [3H]-choline, exogenous NO inhibited electrically-evoked [3H]-choline overflow as well as responses to exogenous agonists, indicating that NO has the potential of neuromodulation both pre- and postjunctionally. 3. A series of NO synthase inhibitors enhanced contractile responses to nerve stimulation indicating inhibitory neuromodulation by endogenous NO. 4. The potency order of the NO synthase inhibitors and their consistent effects after dexamethasone, on responses to nerve stimulation, indicate action on a constitutive NO synthase. 5. Responses enhanced by NO synthase inhibitors were inhibited by the substance P receptor antagonist, spantide, suggesting a neuromodulatory influence on substance P-like neurotransmission by the endogenous NO. 6. NO synthase inhibition did not modify contractile responses to application of acetylcholine or substance P, or [3H]-choline overflow, indicating that endogenous NO mainly has a prejunctional inhibitory action on substance P-like neurotransmission. Nor did it modify responses to direct electrical muscle stimulation in the presence of tetrodotoxin. This suggests a prejunctional enhancing effect by NO synthesis inhibition. 7. Evidence for endogenous NO modulation of acetylcholine release was obtained when NO synthase inhibition modified atropine-sensitive, nerve-mediated contractile responses. However, [3H]-choline overflow was unaltered by NO synthase inhibition. 8. NO synthase inhibition did not modify responses to inhibitory neurotransmission. 9. The findings suggest that endogenous NO inhibits substance P-like motor neurotransmission, probably via prejunctional mechanisms. Cholinergic transmission may also be reduced by endogenous NO, acting prejunctionally.

Modulation of neuroeffector transmission by endogenous nitric oxide: a role for acetylcholine receptor-activated nitric oxide formation, as indicated by measurements of nitric oxide/nitrite release.

Nitric oxide (NO) synthase inhibitors enhanced nerve-mediated contractile responses in guinea pig ileum longitudinal muscle, likely via a prejunctional effect on substance P-like neuroeffector transmission. Supporting a modulatory role for NO, application of NO through administration of acid sodium nitrite evoked marked inhibitory effects on responses to transmural nerve stimulation. Substance P-like responses to nerve stimulation were abolished by substance P receptor antagonists and were enhanced by atropine, indicating a cholinergic influence on substance P-like neuroeffector transmission. Since acetylcholine can evoke release of NO from endothelium, the possible role of acetylcholine in NO release in ileum was examined. The release of NO/nitrite, determined by chemiluminescence, was inhibited by NG-monomethyl-L-arginine (L-NMMA), by calcium removal, by tetrodotoxin or by atropine, indicating a nerve-mediated control of NO production. A basis for the NO release is likely to be spontaneous neuronal activity, where release of acetylcholine, with subsequent muscarinic receptor activation, contributes to stimulation of NO formation.

Endothelin modulation of neuroeffector transmission in smooth muscle.

In a series of muscle preparations, the peptides endothelin-1 (ET-1) and endothelin-3 (ET-3) were investigated for effects on basal muscle tone, responses to transmural nerve stimulation, and release of [3H]norepinephrine or [3H]acetylcholine. ET-1 and ET-3 contracted rat vas deferens and guinea pig ileum, ET-1 being the most potent. In the guinea pig taenia coli, ET-1 induced a relaxation whereas ET-3 was almost without relaxing effect. In the rat vas deferens, ET-1 and ET-3 enhanced contractile responses to nerve stimulation, whereas the nerve-induced release of [3H]norepinephrine was inhibited by ET-1 but not by ET-3. Contractions to exogenous ATP were increased by ET-1 whereas contractions to norepinephrine were not. In the guinea pig ileum, nerve-induced contractions were inhibited by ET-1 and ET-3 as was acetylcholine release, whereas contractions to exogenous acetylcholine were enhanced by ET-1. The inhibition of nerve-induced contractions by the endothelins was not affected by treatment with 8-(p-sulfophenyl)theophylline, BW755C, or indomethacin. The relaxation by ET-1 in the guinea pig taenia coli was not affected by treatment with NG-monomethyl-L-arginine, BW755C, or indomethacin. In conclusion, ET-1 exerted a stimulatory postjunctional effect and concomitantly an inhibitory prejunctional effect on adrenergic and cholinergic neurotransmission. Also, ET-3 exerted a stimulatory postjunctional effect, whereas a prejunctional inhibitory effect of ET-3 only was evident on cholinergic neurotransmission. Blockade of the production of nitric oxide or arachidonic acid metabolites or application of an adenosine antagonist did not alter the effects of ET-1 or ET-3.

Modulation of autonomic neuroeffector transmission by nitric oxide in guinea pig ileum.

NG-monomethyl-L-arginine (L-NMMA), an inhibitor of nitric oxide synthesis, markedly enhanced tonic ("hump") responses to transmural stimulation in guinea pig ileum longitudinal muscle. The enhancement of the hump responses was probably due to a prejunctional effect on substance P-like neurotransmission, since the action of L-NMMA was exerted also in the presence of atropine, and since responses to substance P, a mimic of nerve stimulation, were unaffected by L-NMMA as were cholinergic twitch responses and the overflow of [3H]choline. Further in support, the hump responses were blocked by the substance P antagonist Spantide. All effects of L-NMMA were stereospecifically reversed by L-arginine. Endogenous nitric oxide thus selectively modulates peptidergic neurotransmission in the gut.

Endothelin modulation of neuroeffector transmission in rat and guinea pig vas deferens.

The effects of endothelin-1 (human, porcine) on contractions induced by transmural nerve stimulation, exogenous ATP or noradrenaline, and on the release of [3H]noradrenaline were studied in guinea pig and rat vas deferens. Endothelin enhanced nerve-induced contractile responses, increased basal muscle tone and increased the contractile response to exogenous ATP in both guinea pig and rat vas deferens. Endothelin did not affect the contractile responses to exogenous noradrenaline. The calcium channel blocker felodipine antagonized the stimulating effects of endothelin in the rat vas deferens, whereas blockade of lipoxygenase and cyclooxygenase pathways by a combination of BW 755C and indomethacin was without effect. In rat vas deferens preparations preincubated with [3H]noradrenaline, endothelin inhibited the 3H overflow induced by transmural stimulation, although the contractile responses were enhanced by endothelin. Pretreatment with forskolin or felodipine did not abolish the endothelin inhibition of radiotracer overflow. In conclusion, endothelin can modulate adrenergic and purinergic neuroeffector transmission in both guinea pig and rat vas deferens via inhibitory prejunctional and stimulant postjunctional mechanisms. The stimulant postjunctional effect seemed to predominate in our experiments.

Neuromuscular actions of endothelin on smooth, cardiac and skeletal muscle from guinea-pig, rat and rabbit.

The peptide endothelin (human, porcine) was investigated for effects on basal muscle tone and on responses to transmural nerve stimulation in a series of smooth muscle preparations, as well as in guinea-pig atrium and rat and guinea-pig diaphragm. Endothelin lacked effect on basal tone or on spontaneous and electrically driven contractions in skeletal and atrial muscle. It contracted guinea-pig ileum, pulmonary and femoral arteries, rat anococcygeus, vas deferens and urinary bladder and rabbit taenia coli, whereas guinea-pig taenia was relaxed. Guinea-pig urinary bladder and vas deferens and rabbit iris sphincter were unaffected up to 3 x 10(-8) M. Endothelin thus has a unique pattern of smooth muscle effects, exhibiting mostly contractile but also relaxing effects. Endothelin modified contractile responses to transmural nerve stimulation, yielding marked and persistent enhancement, in guinea-pig and rat vas deferens, and enhancement also in guinea-pig pulmonary artery. In guinea-pig and rat vas deferens the response to exogenous ATP was increased by endothelin, thus suggesting a strong post-junctional enhancement of neurotransmission. In guinea-pig ileum nerve-induced responses were inhibited by endothelin, whereas exogeneous acetylcholine was enhanced, an effect suggesting a simultaneous pre-junctional inhibition and post-junctional enhancement. The Ca2+ channel blocker felodipine counteracted the stimulatory effects of endothelin on tone and transmurally induced contractions. Tachyphylaxis to endothelin action was sometimes evident, but the anococcygeus being less prone to this might be useful for studies on endothelin antagonism. Endothelin thus has prominent post-junctional, and also probably pre-junctional, effects, lending further support for a distinct biological role of this peptide.

Cholinergic neuromodulation by endothelin in guinea pig ileum.

The effect of endothelin on cholinergic neuroeffector transmission in guinea pig ileum was investigated. Endothelin was shown to inhibit the nerve-induced contractions and concomitantly to increase the basal muscle tone. Furthermore, endothelin inhibited the nerve-induced release of [3H]acetylcholine whereas the contractile response to exogenous acetylcholine was enhanced. In conclusion, our findings suggest that endothelin is a modulator of cholinergic neuroeffector transmission in guinea pig ileum with possible action via both inhibitory prejunctional and stimulatory postjunctional mechanisms.