Guinea Pig Lung Parenchymal Strips Research Articles

Electrical Field Stimulation of Isolated Guinea Pig Lung Parenchyma Strips: A Method to Study Peripheral Bronchi and Peripheral Blood Vessels

The reaction of isolated guinea pig lung parenchyma strips to electrical field stimulation (EFS) has been characterized in vitro. Lung parenchyma strips reacted to EFS with a frequency-dependent initial twitch followed by a long-lasting contraction. The initial fast contraction was abolished by tetrodotoxin, whereas the tonic contraction was not influenced by this substance. The initial twitch was reduced but not abolished by atropine as well as by phentolamine. It was concluded that the reaction of guinea pig lung parenchyma strips to EFS consists of a cholinergic and an adrenergic component representing probably the responses of small bronchi and small blood vessels, respectively. The nature of the nonadrenergic-noncholinergic component of the initial twitch as well as of the nature of the nonneural tonic contraction are still unclear.

Lipoxygenase-inhibitory azomethines and benzoylhydrazones. II. Effects of phenylsubstituted azomethines on the antigen-induced contractions of guinea pig lung parenchymal strips and jejunum segments

Lipoxygenase-inhibitory azomethines and benzoylhydrazones. II. Effects of phenylsubstituted azomethines on the antigen-induced contractions of guinea pig lung parenchymal strips and jejunum segments

Fluoride is a contractile agent of guinea pig airway smooth muscle

1. 1. Guinea pig parenchymal lung strips and tracheal smooth muscle contract potently after NaF-addition. Maximal contractions of lung strips and tracheal rings induced by NaF were 208 ± 17% ( n = 6) and 151 ± 8% ( n = 4) of the maximal histamine response respectively. 2. 2. The −log EC 50-value for NaF on lung strips and tracheal rings was 2.38 ± 0.01 ( n = 6) and 2.28 ± 0.01 ( n = 4) respectively. 3. 3. Contractions induced by NaF were augmented after Al 3+ pretreatment, suggesting the involvement of a G-protein. NaF responses were not affected by blockade of H 1-, muscarinic-, leukotriene C 4- or leukotriene D 4-receptors, indicating that mast cell degranulation or nerve activation is most probably not implicated. 4. 4. Contractions after NaF-addition were relatively insensitive to removal of extracellular calcium and were reversed via cAMP- and cGMP-mediated pathways. 5. 5. Relaxation studies with (−)isoprenaline and 8-bromo-cGMP on lung strips, precontracted to similar levels with either a H 1-agonist, KCl or NaF, showed that the level of relaxation depends on the contractile agent that is used. 6. 6. After precontraction with KCl (−)isoprenaline relaxes lung strips only to 58 ± 9% ( n = 5) of the initial contraction, whereas lung strips precontracted with NaF or a H 1-agonist relax 114 ± 8% ( n = 4) and 120 ± 7% ( n = 5) respectively with (−)isoprenaline. 7. 7. Similar results were obtained with relaxation induced with 8-bromo-cGMP. 8. 8. These findings suggest that NaF-induced contractions are elicited via a mechanism, that is probably similar to that of the H 1-receptor. The involvement of a G-protein in the observed NaF-responses is therefore likely.

Essential thiol and disulphide groups in the histamine H1-receptor signal transfer of guinea-pig parenchymal lung strips

Guinea-pig parenchymal lung strips contract after H1-receptor stimulation and membrane depolarisation with KCl. Contractions after 50 mM KCl were similar to the maximal histamine response. Treatment of lung strips with micromolar concentrations of the thiol-alkylator N-ethylmaleimide markedly affects both histamine H1-receptor mediated and 50 mM KCl-induced contractions. The H1-receptor response was only affected via a decrease in the maximal response. The response to 50 mM KCl was also inhibited after thiol-alkylation. However, H1-receptor responses appeared to be slightly more sensitive towards thiol-alkylation compared to KCl-responses. Reduction of disulphide groups with 1,4-dithiothreitol also modified the contractile responses to both stimuli. It is concluded that both thiol- and disulphide moieties play important roles in the regulation of histamine H1-receptor activity.

The Preclinical Pharmacology of ICI 204,219: A Peptide Leukotriene Antagonist

ICI 204,219 (4-(5-cyclopentyloxycarbonylamino-1-methylindol-3-ylmethy l)-3- methoxy-N-o-tolylsulfonylbenzamide) was designed as a peptide leukotriene (LT) antagonist. The compound is a competitive antagonist of LTD4- and LTE4-induced contraction of guinea pig lung tracheal and parenchymal strips with an apparent negative log molar dissociation constant (KB) of approximately 9.6. ICI 204,219 did not antagonize LTC4-induced contractions of guinea pig trachea when the metabolism of LTC4 to LTD4 and, subsequently, to LTE4 was inhibited. The compound inhibited the binding of [3H]LTD4, [3H]LTE4, and [3H]ICI 198,615 (a potent LT antagonist from a different heterocyclic series) to guinea pig lung parenchymal membranes in a competitive manner, and also inhibited [3H]ICI 198,615 binding to human lung parenchymal membranes. ICI 204,219 did not bind to a variety of other receptors when evaluated at concentrations 1,000- to 10,000-fold higher than the apparent KB value for peptide LT receptors. When administered orally, intravenously, or by aerosol, the compound provided dose-related antagonism of the airway effects of aerosol LTD4 in conscious guinea pigs. ED50 values and pharmacodynamic t1/2 (min) for oral, intravenous and aerosol routes of administration were, respectively: 0.52 mumol/kg, greater than 816 min; 0.046 mumol/kg, 85 min; 5.1 x 10(-6) M, 109 min. ICI 204,219 also produced dose-related inhibition of the effects of LTC4 (aerosol or intravenous administration) on pulmonary mechanics in anesthetized guinea pigs when administered orally, intraduodenally, intravenously, or by aerosol. The compound also reversed bronchospasm produced by LTs. Aerosol ovalbumin antigen-induced bronchospasm in guinea pigs was both inhibited and reversed by ICI 204,219. Lastly, the compound inhibited LTD4-induced increases in cutaneous vascular permeability in guinea pigs, being 1,006- and 679-fold more potent than the first generation LT antagonists LY 171,883 and FPL 55712, respectively. ICI 204,219 is a potent, selective, orally active LT antagonist currently undergoing clinical trials.

Ebselen inhibits contractile responses of guinea-pig parenchymal lung strips

Ebselen is a new anti-inflammatory drug with a wide spectrum of pharmacological activities. Since this compound might be useful in diseases related to airway inflammation we evaluated the effects of ebselen on the contractile responses of guinea-pig parenchymal lung strip. Ebselen and its sulfur analogue RP 62373 depressed both histamine H 1-receptor-mediated and KCl-induced (50 mM) contractions of guinea-pig lung strips equipotently. The responses to histamine were only affected via depression of the maximal response; treatment with 3 μM ebselen for 30 min resulted in depression to 77 ± 5% of the control value, whereas 10 and 30 μM inhibited the contractions to 53 ± 4 and 52 ± 4% of the control value respectively. The responses after membrane depolarisation (50 mM KCl) were less sensitive to ebselen pretreatment; 10 μM ebselen inhibited contractions by only 20%, whereas 30 and 100 μM depressed the response by approximately 50%. These observations were evaluated in the context of the activities of ebselen already described. The effects of lipoxygenase, cyclooxygenase, protein kinase C inhibition and thiol alkylation were studied, using established agents. However, although interaction with critical thiol groups might explain our data, the mode of action of ebselen is yet not fully elucidated.

Suppression of PAF-induced bronchoconstriction in the guinea-pig by oxatomide: mechanism of action

Oxatomide potently (ED50 0.9 mg/kg orally, -2 h) attenuates the reduction of pulmonary tidal volume elicited by PAF (250 ng/kg i.v.) in anaesthetized, ventilated and propranolol-treated guinea-pigs. The increase of the pulmonary inflation pressure elicited by PAF (40 ng/kg i.v.) in such animals, ventilated at a fixed tidal volume, is also significantly reduced by the compound, but substantially higher doses (5 mg/kg i.v., -15 min) are required. The potency of oxatomide in the latter respect (50.4% reduction) is equivalent to that of ketotifen at 5 mg/kg i.v. (55% reduction). In spontaneously breathing, anaesthetized guinea-pigs, oxatomide (5 mg/kg i.p., -1 h) significantly reduces the increase in pulmonary resistance, but not the reduction in dynamic compliance, elicited by PAF (30, 60, 90 ng/kg i.v.), suggesting a pharmacological interference mainly with PAF-induced processes in the larger airways. Changes in arterial blood pressure, haemoconcentration, thrombocytopenia and leukopenia induced by PAF in vivo, contraction of guinea-pig lung parenchymal strips, production of superoxide anion by alveolar macrophages, aggregation and release of ATP by platelets challenged with PAF in vitro are not affected by the compound. These observations suggest that oxatomide attenuates the PAF-induced pulmonary reactions by inhibiting the release and/or the effect of allergic mediators elicited by the phospholipid rather than by a direct antagonism at the PAF receptors.

Leukotriene D4 and platelet-activating factor – acether antagonists on allergic and arachidonic acid-induced reactions in guinea pig airways

Arachidonic acid (AA) and ovalbumin (OA) were used to induce contractions of sensitized guinea pig tracheal spiral (indomethacin-pretreated) and lung parenchymal strip preparations. This model was used to examine the properties of three leukotriene (LT) D4 antagonists and a platelet-activating factor (PAF)-acether receptor antagonist. The three LTD4 antagonists, L-649,923, FPL 57231, and LY163443, inhibited AA-induced contractions of indomethacin-pretreated tracheal spirals selectively. The PAF-acether antagonist, L-652,731, did not inhibit AA-induced contractions of either trachea or parenchyma. This confirmed that AA-induced contractions of trachea involved release and activity of LTD4. The LTD4 antagonists and L-652,731 partially inhibited OA-induced contractions of both trachea and parenchyma. When L-649,923 and L-652,731 or FPL 57231 and L-652,731 were combined, an additive inhibitory effect on OA-induced contractions was observed. When LY163443 and L-652,731 were combined, the inhibitory effect was synergistic. This may be due to the additional effect of LY163443 to inhibit phosphodiesterase. Total inhibition of OA-induced contractions was obtainable with relatively low concentrations when a LTD4 and PAF-acether antagonist were combined. These results suggested that LTD4 and PAF-acether may be the two major mediators in our model of allergic bronchospasm. The LTD4 and PAF-acether antagonists had the capacity to decrease baseline tone, even on tissues that were already relaxed with indomethacin, suggesting that LTD4 and PAF-acether may contribute to intrinsic tone in airway smooth muscle.

Influence of platelet-activating factor on leukotriene D4-induced contractions of the guinea pig parenchymal strip.

Platelet-activating factor (PAF) and sulphidopeptide leukotrienes, such as leukotriene D4 (LTD4), are potent constrictors that are probably released simultaneously in a variety of inflammatory respiratory events. The purpose of the present study was to determine whether LTD4-induced contractions of guinea pig parenchymal lung strips (GPPS) are modified in the presence of PAF. The contractile responses of isolated GPPS to cumulative doses of LTD4, acetylcholine, histamine, and potassium chloride in the presence of PAF (0.1 nM, 0.1 microM) were compared with parallel controls. There was no significant alteration of the response to acetylcholine and potassium chloride and the PAF-induced inhibition of the response to histamine, although significant, was not concentration dependent. In contrast, PAF in a concentration range from 0.1 nM to 1.0 microM caused a marked, concentration-dependent reduction of LTD4-induced contractions. Pretreatment with the PAF receptor antagonist, BN52021, prevented the attenuation of LTD4-induced contraction by PAF. The attenuation of LTD4-induced contraction by PAF was also prevented by pretreatment with indomethacin or with the thromboxane synthase inhibitor U63,557A, but not by pretreatment with the lipoxygenase inhibitors BW755c or nordihydroguaiaretic acid. Thus inhibition of LTD4-induced GPPS contraction by PAF is receptor dependent and probably secondary to thromboxane generation. The respiratory smooth muscle response to leukotrienes may be modified significantly by concomitant PAF release.

Biologic activity of a C2-derived peptide. Demonstration of a specific interaction with guinea pig lung tissues.

A synthetic peptide derived from the carboxy terminus of C2b has been investigated for its ability to induce the contraction of guinea pig lung parenchymal strips. This peptide is known to enhance vascular permeability in guinea pig and human skin, and to induce contraction of estrous rat uterus. This C2 peptide (C2 207-223) is active from 5 x 10(-5) M to 5 x 10(-4) M and is not tachyphylactic to itself. No cross-activity between C2 207-223 and C5a or C3a could be demonstrated. C2 207-223 is not inhibited by antihistamines or cyclooxygenase inhibitors. These data indicate that the peptide exerts its action via a mechanism distinct from those of the C3a and C5a anaphylatoxins.

Involvement of protein kinase C in the histamine H1-receptor mediated contraction of guinea-pig lung parenchymal strips.

In this study we investigated the role of protein kinase C (PKC) in the histamine H1-receptor mediated contraction of guinea-pig parenchymal lung strips. Lung strips contract after administration of the PKC activators phorbol-12-myristate-13-acetate (PMA) and dioctanoylglycerol. Prolonged stimulation (45 min.) with a high concentration of PMA abolished subsequent responses to PMA whereas H1-responses were only partially reduced. Administration of a PKC inhibitor (H-7) also led to an inhibition of H1-responses. Experiments with combined incubations with the PKC inhibitor H-7 and the Ca2+-entry blocker verapamil suggest that the PKC contribution to the response is possibly mediated via regulation of Ca2+ influx.

The effect of smooth muscle relaxants working through different transduction mechanisms on the phorbol dibutyrate-induced contraction of the guinea-pig lung parenchymal strip: Possible relevance for asthma

It has been suggested that protein kinase C activation may have a role in the maintained, ‘latch-bridge’ phase of smooth muscle contraction. We have examined the effects of a range of smooth muscle relaxants on the maintained contraction produced in the guinea-pig parenchymal lung strip by the protein kinase C activator, 4β-phorbol dibutyrate (4β-PDBu). The maximum histamine contraction (produced by 10 μM) was used as a standard and the effects of the smooth muscle relaxants were also studied on this histamine-induced contraction. After 4β-PDBu, 1 μM, had produced contraction, enprofylline, forskolin and papaverine caused concentration-dependent relaxation, producing total reversal of the contraction, while prostaglandin E 2 and prostacyclin caused a concentration-dependent relaxation but less than total reversal. The concentrations required for the effects on the phorbol ester contraction were 10 to 100-fold higher than were necessary for relaxation of the maximum contraction produced by histamine. Isoprenaline, 1μM, a concentration which caused total reversal of the histamine-induced contraction, caused only 22% decrease of the phorbol dibutyrate-induced contraction and no further relaxation occurred with higher concentrations. Cromakalim — a potassium channel activator proposed as a therapy for nocturnal asthma — had virtually no effect on preparations pre-contracted with 4β-PDBu, 1 μM, or histamine, 10 μM, but caused about 70% and 20% reversal of the contraction produced by 3 μM histamine and 100 nM 4μ-PDBu respectively. When single doses of the relaxants were administered before a series of doses of 4β-PDBu given cumulatively, enprofylline, 1 μM, and aminophylline, 100 μM and 1 μM, caused a moderate right-shift of the phorbol dibutyrate concentration-response curve, but isoprenaline, 1 μM, was less effective, while cromakalim had no discernible effect. These results are discussed in the light of suggestions that inappropriate activation of protein kinase C in smooth muscle cells, may contribute to the pathogenesis of the late phase of asthma.

Platelet-activating factor-induced contraction of guinea-pig lung parenchymal strips: possible involvement of arachidonate metabolites.

The identification of the mediators possibly involved in platelet-activating factor (PAF)-induced contraction of guinea-pig lung parenchymal strips (GPLP) was attempted by means of antagonists and inhibitors. Histamine, serotonin, acetylcholine (ACh) or other transmitters released from the nerve terminals are not likely to play a role in this response, since specific antagonists and tetrodotoxin did not affect the contraction. PAF antagonists (brotizolam and WEB 2086) produced a concentration-dependent inhibition of the contraction. Inhibitors of TXA2 synthesis (dazoxiben) and of 5-lipoxygenase (nordihydroguaiaretic acid and AA 861) and antagonists of TXA2 (ICI 159995) and peptidoleukotrienes (L 649923 and LY 171883, but not FPL 55712) produced a significant inhibition of the PAF-induced response at concentrations which did not reduce the ACh-induced response. These results suggest that arachidonate metabolites, both of the cyclo-oxygenase and of the lipoxygenase pathway, are determinants of the PAF-induced contraction of GPLP.

The contractile activities of lipoxin A4 and lipoxin B4 for guinea-pig airway tissues.

1. The isometric contractile activities of lipoxin A4 (LxA4) and lipoxin B4 (LxB4) were evaluated on guinea-pig lung tissue over the concentration range, 10(-8) to 10(-5) M. 2. LxA4 contracted guinea-pig lung parenchymal strips; the concentration eliciting 50% maximum histamine response was 3 x 10(-6) M. LxA4 did not contract tracheal spirals. 3. The LxA4 dose-response curve was parallel to that of leukotriene D4 (LTD4) with LxA4 being approximately 10,000 fold less potent than LTD4. 4. The time course of the contraction elicited by LxA4 was similar to that of LTD4 and it was slow in onset and did not plateau for 20 min. 5. Pre-incubation of parenchymal strips with leukotriene receptor antagonists at a concentration of 1 x 10(-6) M to 3 x 10(-5) M FPL 55712 or 3 x 10(-5) M L 649923 inhibited LxA4 activity. 6. Pre-incubation of tissues with 1 x 10(-5) M L 651392, a 5-lipoxygenase inhibitor, or 1 x 10(-5) M indomethacin, a cyclo-oxygenase inhibitor, did not affect the contractile activity of LxA4. 7. LxB4 did not constrict parenchymal strips or tracheal spirals.

Mechanism of action of platelet-activating factor on guinea-pig lung parenchyma strips.

The contribution of thromboxane A2 to platelet-activating factor (PAF)induced contraction of guinea-pig lung parenchyma strips (GPLPS) was investigated using an experimental design that allowed us to record the contractions of the tissues in parallel with the determination of thromboxane B2 (TXB2) levels in the organ baths by enzyme immunoassay. It was found that the first injection of PAF induced the contraction of GPLPS and the release of TXB2. Following subsequent additions of PAF to the same tissue, the contractile response was abolished but TXB2 levels were not significantly reduced. Pretreatment of the tissue with the thromboxane synthetase inhibitor OKY-046 (3.5, 170, and 350 microM) strongly inhibited the release of TXB2 but had no effect on the contraction of the tissues induced by PAF. The mechanism of PAF-induced contraction of GPLPS was further investigated using several drugs that interfere with arachidonic acid metabolism. It was found that pretreatment of the tissues with the cyclooxygenase and thromboxane synthetase inhibitors indomethacin (2.8, 28, and 56 microM) and OKY-046 (170 microM) or with the thromboxane antagonist SKF-88046 (1.25 and 12.5 microM) had no significant effect on the contractile response to PAF. The compound L-655,240 (2.5, 25, and 50 microM), which acts simultaneously as an antagonist of thromboxane and inhibitor of lipoxygenase, significantly reduced GPLPS contractions induced by PAF. Another lipoxygenase inhibitor, nordihydroguaiaretic acid (33 microM), and the inhibitor of both pathways of arachidonic acid metabolism, BW775c (110 microM), both reduced PAF-induced contractions of GPLPS.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of LG 30435 on different platelet activating factor-induced responses.

LG 30435 is a new potential anti-asthmatic agent with multiple sites of action. It can inhibit the bronchoconstriction induced by various agonists, including platelet activating factor (PAF). The effects of this compound on PAF-induced biological responses have now been investigated. LG 30435 inhibited both serotonin release from, and aggregation of, rabbit washed platelets evoked by PAF, being four times more potent on release than on aggregation. In contrast, it did not inhibit a number of the biological effects of PAF which are not platelet-mediated, such as contraction of guinea-pig lung parenchymal strips, hypotension in rats and lethality in mice. It may be concluded that, unless platelet PAF receptors are different from those in other tissues, LG 30435 is capable of inhibiting platelet-mediated effects of PAF by interfering with secondary mechanisms activated by this autacoid and not by direct antagonism of PAF at receptor sites.

A formyl peptide contracts guinea pig lung: role of arachidonic acid metabolites.

We studied the role of cyclooxygenase and lipoxygenase products of arachidonic acid metabolism in mediating N-formyl-methionyl-leucyl-phenylalanine- (FMLP) induced contractions of guinea pig lung parenchymal strips. The cyclooxygenase inhibitors indomethacin (10(-5) M) and aspirin (3 X 10(-5) to 10(-4) M), the lipoxygenase inhibitor nordihydroguaiaretic acid (10(-5) to 3 X 10(-5) M), and the combined cyclooxygenase/lipoxygenase inhibitors 1-phenyl-3-pyrazolidinone (Phenidone) (3 X 10(-5) to 3 X 10(-4) M) and BW 755C (10(-5) to 10(-4) M) each caused a decrease in the maximum force induced by FMLP (Fmax) and an increase in the concentration of FMLP required to produce 50% of Fmax (EC50). The thromboxane synthesis inhibitor imidazole (3 X 10(-3) M) also decreased Fmax. The leukotriene D4 receptor antagonist FPL 55712 (5.7 X 10(-6) to 1.9 X 10(-5) M) increased the EC50 for FMLP, whereas desensitization of lung parenchymal strips to leukotriene B4 by pretreatment with this leukotriene (10(-7) M) had no effect on FMLP-induced contraction. After exposure to FMLP (10(-6) M), guinea pig lung produced (as determined by high-performance liquid chromatography and radioimmunoassay) leukotrienes C4 and B4, thromboxane A2 (as measured by its stable degradation product thromboxane B2), and prostaglandin F2 alpha. Lung strips not exposed to FMLP showed no evidence of leukotriene production. We conclude that thromboxane A2 and leukotriene C4 generated in response to FMLP mediate a substantial fraction of the force induced by this peptide in guinea pig lung parenchymal strips.

Effects of gossypol on guinea pig airway responsiveness: in vitro and in vivo studies

AbstractThe actions of the naturally occurring polyphenol, gossypol, on guinea pig lung parenchymal strip preparations, bronchoconstriction in anaesthetized guinea pigs and platelet aggregation were examined. At concentrations between 0.2 and 20 μg/ml gossypol markedly inhibited contraction of lung parenchymal strips induced by LTB4 and LTD4, but was less effective against histamine‐induced contraction. Gossypol also inhibited contraction induced by platelet‐activating factor‐acether (PAF‐acether) but was not as effective as BN52021, a specific PAF‐acether antagonist. In addition, gossypol failed to antagonize PAF‐acether‐induced bronchoconstriction in anaesthetized guinea pigs or the histamine‐dependent bronchoconstriction response to ovalbumin in passively sensitized animals. Gossypol was minimally effective in inhibiting PAF‐acether‐induced platelet aggregation. This polyphenol displays a similar spectrum of effects to the radical scavenger NGDA, suggesting that, in common with other phenolic antioxidants, it may interfere with the formation of cyclo‐oxygenase products in the guinea pig lung.

Neutral endopeptidase-like enzyme controls the contractile activity of substance P in guinea pig lung.

The responsiveness of isolated guinea pig lung parenchymal strips to substance P was enhanced by at least 100-fold in the presence of the endopeptidase inhibitors phosphoramidon (1 microM) or thiorphan (1 microM), but not with the converting enzyme inhibitor, captopril, or an inhibitor of serum carboxypeptidase N (both 1 microM). Responses of guinea pig tracheal rings to substance P were also markedly potentiated by phosphoramidon. The increase in tissue responsiveness by these inhibitors was relatively specific for substance P among several other spasmogenic peptides, including formyl-methionyl-leucyl-phenylalanine and the complement peptides C3a and C5a. The enhanced responses appear to result from a decrease in the rate of substance P degradation in the presence of neutral endopeptidase inhibitors. Specific binding of substance P to its receptor on bronchial membranes was increased by three- to fourfold in the presence of phosphoramidon. These data demonstrate an enhanced potential for substance P to contract lung tissues when degradation by a neutral endopeptidase-like enzyme is blocked.

Modulation of the contractile activity of the guinea-pig lung parenchymal strip by exogenous 5,8,11,14,17-eicosapentaenoic acid.

Exogenous eicosapentaenoic acid (EPA, 16.5 mumol/l or 33 mumol/l) inhibited dose-dependently the anaphylactic contractile response of guinea-pig lung parenchymal strips suspended in an organ bath. As determined by radioimmunoassay, EPA inhibited in a dose-dependent manner the anaphylactic release of the cyclooxygenase products thromboxane (TX) B2 and 6-keto-prostaglandin (PG) F1 alpha but simultaneously enhanced the release of sulfidopeptide (SP)-leukotrienes (LT). Indomethacin (2.8 mumol/l) abolished the release of cyclooxygenase products but potentiated the release of SP-LT. However, indomethacin treatment did not affect the inhibitory action of EPA on the contractile response of the anaphylactic lung strips. The lipoxygenase inhibitor, esculetin (50 mumol/l), inhibited the release of SP-LT and also that of cyclooxygenase products of polyunsaturated fatty acid metabolism. The combination of esculetin and EPA resulted in enhanced inhibition of the anaphylactic contractile response as compared to EPA alone. By reversed phase high pressure liquid chromatography (HPLC), SP-LT from anaphylactic lung parenchymal strips was shown to consist of LTD4 and LTE4. EPA-pretreated lung strips released upon immunologic challenge additional immunoreactivity comigrating with authentic LTC4, LTC5, LTD5 and LTE5. While anaphylactic control strips also released LTB4, in the bath fluid of EPA-treated strips, an additional immunoreactive compound migrating with the retention time of LTB5 was observed. In non-sensitized guinea-pig lung parenchymal strips EPA inhibited the myotropic activity of exogenous mediators such as histamine (9 mumol/l), LTC4 (16 nmol/l) and the TX mimetic U 46619 (28.4 nmol/l), an effect which was neither affected by indomethacin (2.8 mumol/l) nor by the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA, 10 mumol/l).(ABSTRACT TRUNCATED AT 250 WORDS)