Cyclooxygenase mediated airway response to leukotriene D4 in the cat

The effects of leukotriene D₄ (LTD₄) on pulmonary mechanics were investigated in anesthetized, paralyzed cats under conditions of controlled ventilation. Intravenous injections of LTD₄ in doses of 3, 10, and 30 μg caused significant increases in transpulmonary pressure (P_TP) and lung resistance (R_L) while decreasing dynamic compliance (C_dyn). LTD₄ also increased systemic arterial pressure (P_A0). The changes in P_TP, R_L, and C_dyn in response to LTD₄ were blocked by sodium meclofenamate, a cyclooxygenase inhibitor. However, there was no significant change in the increase in P_A0 following cyclooxygenase blockade. U 46619, a thromboxane mimic, was 30 to 100 times more potent than LTD₄ in increasing P_TP, R_L and decreasing C_dyn in the cat. These data show that LTD₄ has significant smooth muscle constrictor activity in central airways as well as peripheral portions of the feline lung. In addition, these data suggest that in the cat the actions of intravenously administered LTD₄ on lung mechanics are mediated by release of cyclooxygenase products while the systemic pressor effects are not dependent upon the integrity of the cyclooxygenase pathway.     

Pharmacology of peptide leukotrienes on ferret isolated airway smooth muscle

The contractile activities of peptide leukotrienes (LT) on isolated spiral strips of ferret trachea were chracterized pharmacologically. LTC₄ and LTD₄ contracted ferret tracheal strips in a concentration-related manner and were 3- to 8-fold more potent than carbachol. In contrast, high concentrations of LTE₄ evoked either weak contraction or none at all, whereas LTC₄ and D₄ were partial agonists compared to carbachol. In tissues which were unresponsive to LTE₄, this compound antagonized contractile responses to LTC₄ and D₄ in an apparently competitive manner: Carbachol-induced contractions were not altered by LTE₄. The cyclooxygenase inhibitor, indomethacin (5 μM), LT antagonists, FPL55712 (10 μM), atropine (1 μM), phenoxybenzamine (10 μM), and LTB₄ (10 μM) failed to alter LTC₄ and D₄ concentration-response curves. The results in dicate that ferret trachea is sensitive to the contractile activity of LTC₄ and LTD₄ but not LTE₄. The LT-induced contractions appear to be mediated by a direct action of the LT rather than indirectly through release of secondary mediators such as thromboxane, prostaglandin, or acetylcholine. LT receptors in ferret trachea are insensitive to FPL55712 but are antagonized by LTE₄.     

Antagonism by ETYA of the effects of leukotrienes on ileum and lung parenchymal strips independent of effects on arachidonic acid metabolism

5,8,11,14-Eicosatetraynoic acid (ETYA), a compound which inhibits both the cyclooxygenase and lipoxygenase pathways of arachidonic acid metabolism, antagonized the contraction of segments of guinea-pig ileal longitudinal muscle produced by SRS-A (IC₅₀ = 2.73 μM). This activity was unaffected by pretreatment of the tissues with 10 μM indomethacin. Phenidone, another mixed cyclooxgenese-lipoxygenese inhibitor, was inactive. FPL-55712, an SRS-A antagonist, was a very potent inhibitor (IC₅₀ = 0.011 μM).BW755C and NDGA nonselectively inhibited the contractions of the guinea-pig ileal longitudinal muscle induced by SRS-A or histamine.ETYA antagonized the contraction of the guinea-pig ileal strip produced by 6 nM synthetic LTC₄ (IC₅₀ = 9.3 μM). FPL-55712 demonstrated an IC₅₀ of 0.3 μM in a similar series of experiments. ETYA, 1, 3 or 10 μM did not inhibit the contractions elicited by 0.5 μM of histamine.This was not a tissue-selective effect since 100 μM ETYA antagonized the LTC₄-induced contraction of the guinea-pig lung parenchymal strip preparation.These data demonstrate that ETYA antagonized the contractile effect of the leukotrienes on tissues from the gastrointestinal tract and lung. Furthermore, the inability of indomethacin or phenidone to inhibit the contractile response suggests that antagonism by ETYA may occur by a mechanism independent of cyclooxygenase and lipoxygenase enzymes.     

Evidence for a mediator role of thromboxane A2 in the myotropic action of leukotriene B4 (LTB4) on the guinea-pig lung

The mechanism of action of LTB₄ has been investigated on the guinea-pig lung parenchymal strip. Mepacrine (20 ug/ml), an inhibitor of phospholipase A₂, abolished the action of LTB₄ on parenchymal strips. Eicosatetraynoic acid (10 ug/ml) and BW755C (40 ug/ml) which are inhibitors of cyclooxygenase and lipoxygenase pathways, produced a marked inhibition of the lung strip contraction to LTB₄. Similarly, aspirin (30 ug/ml) and flufenamate (lug/ml) showed a strong inhibition of the contraction of parenchymal strips to LTB₄; these results suggested that cyclooxygenase products mediate the action of LTB₄. The response to LTB₄ was unaffected by 15-hydroperoxyeicosatatraenoic acid (15-HPETE; 1 ug/ml) while L8027 (25 ng/ml) reduced the contraction by 50%, suggesting that thromboxane A₂ rather than prostacyclin was involved. Since parenchymal strips do not appear to be very sensitive to PGF₂α, PGE₂ and the endoperoxides, and since effluents from LTB₄-treated lungs produced contractions of lung strip and rabbit aorta which were reduced after 5 min. at 250, thromboxane A₂ was postulated to mediate the lung effect of LTB₄. The release of thromboxane B₂ (TxB₂) from lungs stimulated with LTB₄ was confirmed by gaschromatography-mass spectrometric (GC-MS) analyses.     

Differing mechanisms for leukotriene D4-induced bronchoconstriction in guinea pigs following intravenous and aerosol administration

Leukotriene D₄ (LTD₄) when administered intravenously or by aerosol to guinea pigs produced changes in pulmonary mechanics including a decrease in dynamic compliance and an increase in pulmonary resistance. The effects of intravenous LTD₄ (0.5 μg kg⁻¹) were short lived and abolished by pretreatment of the animal with either cyclooxygenase inhibitors, a thromboxane synthetase inhibitor (OKY 1555) or an SRS-A antagonist (FPL 55712). These findings suggest that bronchoconstriction produced by the intravenous infusion of LTD₄ at 0.5 μg kg⁻¹ is due to the release of thromboxane A₂. However, in animals treated with indomethacin, LTD₄ at higher doses (>0.8 μg kg⁻¹) still elicited a bronchoconstriction which could be blocked by FPL 557112. Nebulization of 0.1 – 1.0 μg of LTD₄ into the lung produced prolonged changes in pulmonary mechanics which were inhibited by FPL 55712 and were potentiated indomethacin. LTD₄, therefore, when administered by aerosol produced effects on the lung which were not mediated by cyclooxygenase products. Responses to nebulized rather than intravenous LTD₄ in the guinea pig may more closely resemble those seen in human tissues.     

A proposed role for prostaglandins in the modulation of the relaxation response to urotensin I in isolated rat arteries

Urotensin I (UI) elicits dose-dependent relaxation responses in isolated helical strips of rat tail and mesenteric arteries contracted by 10⁻⁵M norepinephrine (NE). The rat mesenteric artery demonstrated a 40 fold lower threshold sensitivity to UI (0.25 mU/M1 versus maximal relaxation at 0.25 mU/m1). Complete relaxation of the rat tail artery with UI could not be achieved, even at doses exceeding 10 mU/m1. Pretreatment of the arterial strips with cyclooxygenase inhibitors had no effect on the contractile response to NE in the tail artery, but reduced NE responsiveness in the mesenteric artery. Significant enhancement of UI relaxation responses in both types of arterial strips was achieved by pre-treatment with the cyclooxygenase inhibiters, suggesting a modulatory role for prostaglandins (PGs) in the expression of the UI relaxation response in NE contracted arterial strips. The major enzymatically formed PG (as assessed by [1-¹⁴C] PGH₂ metabolism in broken cell preparations) in both the rat tail and mesenteric arteries was 6-keto PGF_1α, the stable hydrolysis product of PGI₂. Using a specific RIA to quantify 6-keto PGF_1α release, it was found that UI elicited nearly a two-fold increase in the release of this PG compared to the NE control in both rat tail and mesenteric arteries. These data suggest that PGI₂ may modulate the relaxation response to UI either by direct physiological opposition (PGI₂ elicited contractile response in NE contracted tail and mesenteric arteries at doses exceeding 10−8M) and/or by some as yet undefined mechanism (eg. effects on Ca²⁺, cAMP).     

Inhibition of pulmonary thromboxane A2 synthase activity and airway responses by CGS 13080

The effects of CGS 13080, a thromboxane (TXA₂) synthase inhibitor, on airway responses to arachidonic acid (AA) were investigated in the anesthetized cat. Feline and human lung microsomal fraction exhibited prostaglandin I₂ (PGI₂, prostacyclin), and TXA₂ synthase activities, and human platelet microsomal fractions exhibited TXA₂ synthase activity. Cat and human lung microsomal fractions, but not human platelets, exhibited the presence of GSH-dependent PGE₂ isomerase activity. CGS 13080 inhibited TXA₂ synthase activity in all three microsomal fractions in a concentration-dependent manner. The increases in transpulmonary pressure and lung resistance and decreases in dynamic compliance in response to AA were decreased significantly by CGS 13080. These data suggest that the bronchoconstrictor actions of AA are mediated in large part by the formation of TXA₂. The data further indicate that cyclooxygenase products other than TXA₂ are involved in the bronchoconstrictor response to AA since meclofenamate had greater inhibitory activity than did CGS 13080. Moreover, the effects of CGS 13080 were due to inhibition of TXA₂ synthase rather than an effect on TXA₂ receptors, since airway responses to the TXA₂ mimic, U46619, were not altered. The present data show that CGS 13080 inhibits TXA₂ synthase activity without altering cyclooxygenase, PGI₂ synthase, or GSH-dependent PGE₂ isomerase activities. The data further indicate that in vivo administration of CGS 13080 may selectively increase PGI₂ synthase activity.     

Interaction between the M2- and M3-Receptor Subtypes in Muscarinic Electrical and Mechanical Responses of Intestinal Smooth Muscles

In various gastrointestinal smooth muscles, two different muscarinic receptor subtypes, M₂ and M₃, are expressed; these receptors are the target for the parasympathetic neurotransmitter acetylcholine. Although the number of M₂ receptors is much greater than that of M₃ receptors, the functional role of the former receptor subtype has yet to be fully defined, since pharmacological analyses of the contractile responses to acetylcholine and other muscarinic agonists have revealed that such responses are mediated extensively by the minor M₃ subtype. The M₃ receptor links to Ca²⁺ store release, and the released Ca²⁺ ions may contribute to the contraction. However, many studies indicated the importance of Ca²⁺ influx through voltage-gated Ca²⁺ channels, rather than Ca²⁺ release, in muscarinic contractions, since the contractile responses are markedly inhibited by Ca²⁺ channel blockers. The major M₂ receptors link to the opening of cationic channels leading to the membrane depolarization, which in turn activates voltage-gated Ca²⁺ channels. Thus, there should be somewhere a point of contact between the M₃- and M₂-mediated signal transductions, as if M₃ receptor stimulation is connected with membrane depolarization. Our electrophysiological and pharmacological findings suggest that the M₂-mediated cationic channel opening and a resulting increase in the membrane electrical activity are the primary mechanism for mediating the contractile response to muscarinic agonists. An allosteric interaction between M₂ and M₃ receptors such that M₃ activation intensifies the M₂/cation channel pathway may account at least in part for the failure of many previous analyses to detect M₂ participation in the contractile responses to full agonists.     

Further studies on the mechanism of action of leukotrienes and histamine on guinea pig lung parenchyma role of calcium, phospholipase and methyltransferase

The contractile activity of leukotriene B₄ (LTB₄), leukotriene D₄ (LTD₄) and histamine on strips of guinea pig lung parenchyma was shown to be dependent on the calcium concentrations of the Krebs solution. The calcium channel blocker verapamil (2.0 to 15uM) had an additive effect on the inhibitory activity of low calcium (0.1 mM) on contractions of guinea pig parenchyma to leukotrienes and histamine. Cobalt chloride, a divalent cation, also produced dose-dependent reductions of the myotropic activities of LTB₄, LTD₄ and histamine. An antagonist of calmodulin, triflouperazine (1–200 uM), dose-dependently inhibited the contractile activity of the three agonists on the parenchyma strip. The IC₅₀ of this compound for inhibition of histamine was much lower (2–3uM) than the IC₅₀ for inhibition of leukotrienes (75 uM). Valinomycin, a potassium ionophore, also interfere with the contractile activities of leukotrienes and histamine whereas a blocker of sodium channel, tetrodotoxin, had no effect on the activity of these agonists. Furthermore, an inhibitor of methyltransferase, 3-deazaadenosine, significantly diminished the responses of the parenchyma to leukotrienes and histamine. These results confirmed the important role of extracellular and intracellular calcium in the myotropic activity of leukotrienes and histamine in guinea pig lungs and showed that compunds which interfere either directly or indirectly with calcium mobilization into the lung smooth muscles, decreased the tissue responsiveness.     

Leukotriene F4 and the release of arachidonic acid metabolites from perfused guinea pig lungs in vitro

Radioimmunoassay and bioassay techniques have been used to investigate the ability of leukortriene (LT)F₄ to release products of arachidonic acid metabolism from guinea pig isolated lungs perfused via the pulmonary artery. Also, the abilities of LTC₄, LTD₄ LTE₄ and LTE₄ to contract guinea pig ileal smooth muscle (GPISM) was studied. Each of the LT's contracted GPISM. The rank order of potency was LTD₄ > LTC₄ > LTE₄ > > LTF₄ in a ratio 1:7:170:280 respectively. Bioassay of pulmonary effluents indicated the passage of LTF₄ through the lungs caused a contraction of rabbit aorta as well as an FPL-55712 sensitive contraction of GPISM. The contractions of rabbit aorta were inhibited by pretreatment of the lungs with Indomethacin but not with the thromboxane synthetase inhibitor Dazoxiben. Radioimmunoassay of the lung effluents indicated LTF₄ to cause a 70-fold increase in thromboxane B₂ (TXB₂), 4-fold increase in prostaglandin (PG)E₂ and a 16-fold increase in 6-keto PGF_1α levels. The LTF₄-induced increments of these immunoreactive metabolites was inhibited by pretreatment of the lungs with Indomethacin. Pretreatment of lungs with Dazoxiben inhibited the LTF₄-induced increment in TXB₂ and enhanced the effluet levels of PGE₂ 24-fold (compared with untreated lungs). There were no detectable differences in either immunoreactive LTC₄ or immunoreactive LTB₄ levels. It is concluded LTF₄ is a relatively weak agonist on GPISM and can induce the release of cyclooxygenase products of arachidonic acid metabolism from guinea pig perfused lung.     

Quantitation of luekotrienes C4, D4 amd E4 released by pathophysiological stimuli

In order to examine the modulation of leukotriene (LT) release, the PAF-acether-mediated stimulation of these compounds in rat lung was studied. Release of LTC₄, LTD₄ and LTE₄ in both perfused and chopped lung preparations was measured using HPLC and radioimmunoassay. Pre-incubation or pre-infusion of the tissue with indomethacin and PGE₂ was conducted to investigate the effect of cyclooxygenase inhibitors and products on the lipoxygenase pathway. In addition, the effects of LT levels of pre-incubation with vasoactive intenstinal polypeptide (VIP) in chopped lung were observed.In perfused rat lung, indomethacin reduced the levels of LTC₄ relative to LTD₄ as measured in the first 2 min after stimulation of the lung by PAF-acether. Chopped lung preparations, incubated for 15 min. exhibited higher levels of LTC₄ and LTD₄ in indomethacin-treated samples, this increases being effectively reversed by PGE₂.In the VIP pre-incubation experiments clear inhibition of peptido -leukotriene synthesis was observed, with no LTC₄ and only low levels of LTD₄ and LTE₄ observed in VIP-incubated samples. In preliminary experiments using rabbit C5a des arg and PAF-acether on rabbit lung parenchyma strips to stimulaet LT release, disodium cromoglycate pre-incubation was observed to inhibit this release.Inhibition of the 5-lipoxygenase pathway of PGE₂ is supported by these experiments. VIP appears to act as an inhibitor of LTC₄ and LTD₄ biosynthesis or release in this model. Too little is known that peptidergic actions to postulate a mechanism by which a neuroendocrine peptide exerts control of release of arachidonate metabolites; however, VIP is associated with muscarinic stimulation (1) and has been found in mast cells (2).     

Contraction of guinea pig gail bladder strips by leukotrienes and other agonists

In the presence of indomethacin, Leukotriene C₄ (LTC₄), LTD₄ and LTE₄ were shown to be contractile agents on guinea pig gall bladder strips. The respective pD₂ values for LTC₄, LTD₄ ad LTE₄ were 9.1, 9.1 and 7.7. The contractile effects of LTD₄ were not mediated through the generation of cyclooxygenase products and were antagonized by the SRS-A antagonist FPL-55712. The effects of PGE₁, PGF_2α, the endoperoxide analogue U44069 and histamine on gall bladder strips were also examined. All these agents caused dose-related contractions but were considerably less potent than the leukotrienes. Leukotrienes are therefore potent contractile agents on the guinea pig gall bladder and may contribute to gall bladder contractions or spasms .     

Effects of pure enantiomers of flurbiprofen in comparison to racemic flurbiprofen on eicosanoid release from various rat organs ex vivo.

The effects of oral treatment of rats with pure enantiomers of flurbiprofen in comparison to racemic flurbiprofen on ex vivo release of eicosanoids from gastric mucosa, jejunum, lung, brain and clotting whole blood were investigated. With the S(+) enantiomer and the racemate dose-dependent inhibition of release of cyclooxygenase products of arachidonate metabolism in all tissues tested was observed, while release of leukotriene (LT) C4 was inhibited in gastric mucosa, but not in jejunum and lung. On the other hand, the R(-) enantiomer inhibited cyclooxygenase in the various tissues less potently and to a variable degree with no significant effect in the jejunum. The R(-) enantiomer had no effect on LTC4 release from any of the tissues investigated. Furthermore, the effect of a high dose of 25 mg/kg of the S(+) enantiomer on release of cyclooxygenase products from the various tissues was much longer lasting than that of an identical dose of the R(-) enantiomer. Stereoselective pharmacokinetics of the flurbiprofen enantiomers and/or organ specific cyclooxygenase activities could underly these results. The more potent cyclooxygenase inhibition by the S(+) enantiomer correlates with its higher anti-inflammatory activity and gastrointestinal toxicity. On the other hand, both enantiomers have been shown previously to be almost equally effective analgesics. Inhibition of brain cyclooxygenase might contribute to this effect.     

Induction of Prostanoid Biosynthesis by Bacterial Lipopolysaccharide and Isoproterenol in Rat Microglial Cultures

Abstract: We have used purified microglial cultures obtained from neonatal rat cerebral cortex to investigate the ability of microglia to release prostanoids after exposure to bacterial lipopolysaccharide, a classic macrophage activator. Release of prostaglandin E₂, prostaglandin D₂, and thromboxane A₂ was low in basal conditions and increased in a dose- and time-dependent way upon lipopolysaccharide treatment (1–100 ng/ml), by a mechanism requiring de novo protein synthesis. When compared with astrocytes, microglial cells appeared to respond more effectively to lipopolysaccharide, being able to release prostanoids after exposure to a 100-fold lower concentration of lipopolysaccharide. In addition to prostanoids, we also measured the release of leukotriene B₄; although lipopolysaccharide failed to stimulate leukotriene B₄ release by microglial cells, it doubled the basal production in astrocytes. Lipopolysaccharide enhanced the release of preloaded [³H]arachidonic acid from microglial membrane phospholipids by a mechanism inhibited by the protein synthesis inhibitor cycloheximide, which suggests that the increased availability of arachidonic acid contributed to the enhanced prostanoid production. Lipopolysaccharide, however, also stimulated prostanoid synthesis by inducing cyclooxygenase activity, as shown by determining the activity of newly synthesized enzyme after inactivating the endogenous enzyme with aspirin and by assessing the level of the inducible form of cyclooxygenase by western blot analysis. Among the mechanisms potentially involved in the regulation of microglial prostanoid production, we studied the effect of β-adrenergic receptor activation. The β-agonist isoproterenol was inactive by itself but doubled the effect of lipopolysaccharide. The drug appeared to act mainly through the inducible cyclooxygenase; because it did not stimulate arachidonic acid release, it enhanced the lipopolysaccharide-evoked prostanoid production observed after aspirin pretreatment and induced de novo synthesis of cyclooxygenase detectable by western blot analysis. We suggest that during cerebral inflammatory processes microglia can contribute to the establishment of high prostanoid levels, which can be further elevated by β-adrenergic activation.     

An inhibitory effect of arachidonic acid on subsequent responses of canine cerebral arteries to serotonin and other agonists

Arachidonic acid (AA) at 10^?4M and 10^?3M produced a phasic contraction in isolated canine basilar arteries that peaked rapidly and then slowly declined. This contraction was evidently due to the conversion of AA to prostanoids because it was blocked by cyclooxygenase inhibitors and because 11, 14, 17 eicosatrienoic acid (10^?3M), which is not a cyclooxygenase substrate, failed to produce a contraction. When the artery was exposed to 10^?3M AA for 20 min and washed, subsequent contractile responses to 10^?6M serotonin (5-HT) were only 10% of control. Contractions produced by prostaglandin E₂ (10^?5M), uridine triphosphate (10^?4M) and potassium (5.5×10^?4M) were inhibited to a lesser degree than 5-HT, the response to potassium being the least affected (66% of control). This damaging effect of 10^?3M AA did not occur if the artery was washed at peak contraction nor with 10^?4M AA. Autooxidation products were evidently not responsible for the damage because prior oxygenation (90 min) of 10^?4M AA had no such effect. Pretreatment with superoxide dismutase or ascorbate did not prevent the inhibition, suggesting that free radical reactions were not involved. Pretreatment with indomethacin (3×10^?4M) or meclofenamate (10^?4M) also failed to prevent the inhibitory phenomenon. Saponin, a detergent, produced similar inhibitory effects but 11, 14, 17 eicosatrienoic acid or oleate (10^?3M) did not. The arteries partially recovered from the inhibition with time. In conclusion, AA produced contraction in basilar arteries by inducing prostaglandin synthesis but can produce secondarily by an unidentified mechanism an inhibition of the contractile responses evoked by various agonists that is both time and concentration dependent.     

The effect of calcium antagonists on contractions of the sensitized and normal guinea pig ileum

The purpose of this study was to learn wether a number of Ca²⁺ antagonists were effective in reducing contractile response of the isolated ileum of the sensitized and normal guinea pig. Contractions of the normal ileum in response to LTD₄, acetylcholine, histamine, and potassium chloride were obtained before and after verapamil, diltiazen and papaverine. Ovalbumin-induced contractions of the ovalbumin-sensitized ileum were obtained in the presence of the three Ca²⁺ antagonists. In the normal ileum, all the Ca²⁺ antagonists were highly effective in diminishing the contractile responses to LTD₄, acetylcholine, histamine and potassium chloride. In the sensitized ileum, ovalbumin-evoked contractions, with subsequent release of a potent contractile mediator (presumably SRS-A), were Ca²⁺-dependent since verapamil, diltiazem and papaverine caused a concentration-related reduction of contractions. Thus, the influx of extracellular Ca²⁺ plays a key role in the contractile responses of the normal and sensitized guinea pig ileum when stimulated by various potent agonists acting on specific receptors or on the cell membrane.     

Histamine releases PGI2 from human pulmonary artery

Histamine caused a triphasic response of human pulmonary artery strips in vitro, consisting of a small initial contraction followed by pronounced relaxation preceding a second contractile response. These characteristics were not seen with other contractile stimuli including 5-hdyroxytryptamine, leukotriene D₄, and KC1. The relaxant component of this response was ablated by removal of endothelium from the vascular strips or by pretreatment of the tissues with 1μM indomethacin. Measurement of the PGI₂ degradation product 6-keto-PGF_1α in supernatants from histamine-challenged tissues confirmed the synthesis of PGI₂. Supernatants from unstimulated or leukotriene-challenged tissues contained no detectable amounts of 6-keto-PGF_1α. The histamine H₁ antagonist diphenhydramine inhibited both the contractile and relaxant responses to histamine whereas the H₂ antagonist cimetidine affected neither component. The released PGI₂ significantly altered the dose-respons curve to histamine without inhibiting the maximal contractile responses. We conclude that histamine induces PGI₂ formation from pulmonary arterial endothelium via an H₁ receptor.     

Transformation of individual contractile responses during tetanus in rat fast and slow skeletal muscles

Using a computer graphics approach, the last contractile responses (LCR_N, where N is a number of elementary contractile responses in tetanus) were separated from integral tetanic responses of rat fast muscles, m. Eхtensor digitorum longus (m. EDL), and slow muscles, m. Soleus, evoked by trains of 5, 10 and 50 stimuli. In m. Soleus, at a stimulation frequency of 20 Hz, the LCR₅ average amplitude decreased to 64 ± 9% compared to the single contraction amplitude. As N increased, LCR_N recovered and then rose to the values exceeding almost twofold initial elementary contractile responses (up to 211 ± 10% for LCR₅₀). Simultaneously, against the background of rising elementary contractile responses, a significant shortening of their half-decay time (～by 50%) and the formation of a stationary plateau within LCR_N was observed. In m. EDL, at a stimulation frequency of 50 Hz, there was only a single-phase LCR_N rise (up to 165 ± 18% for LCR₅₀) without changes in half-decay time and plateau formation. In skeletal muscles of both types, the prolonged (up to 30 s) ‘hyper-relaxation effect’ was found to develop after the end of tetanic responses manifested as a reduction of muscle tension followed by its recovery to the initial level. Possible mechanisms of these events are discussed. It is hypothesized that transformation of elementary contractile responses in skeletal muscles can be fulfilled due to the existence of specialized microdomains in muscle fibers which regulate accumulation and extrusion of Ca²⁺ ions during tetanic activity. The possibility that the basic, depolarization-induced, Ca²⁺ release (DICR) is complemented by an additional, Ca²⁺-induced, Ca²⁺release (CIRC) is analyzed.     

Bornchoconstriction induced by N-formyl-methionyl-leucyl-phenylalanine in t he guinea pig; involvement of arachidonic acid metabolites

The chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (F-met-leu-phe) was shown to be a potent myotropic agent on the guinea-pig parenchymal strip (IC₅₀), 2 × 10⁻⁷ M). The response was unaffected by the histamine (H₁) antagonist, mepyramine, but in the presence of the cyclooxygenease inhibitor, indomethacin, the dose response curve was shifted to the left (IC₅, 4 × qo⁻⁸ M) and the maximal response reduced. Injection of F-met-leu-phe into perfused guinea pig lungs resulted in the release of leukotriene-like activity which was detected by superfusion over guinea-pig ileum preparations in the presence and absence of FPL-55712. Intravenous injection of F-met-leu-phe to spontaneously breathing anaesthetized guinea pigs resulted in transient increases in pulmonary resistance and blood pressure and decreases in dynamic compliance and heart rate. The pulmonary parameters were significantly inhibited by BW 755C, indomethacin, FPL-55712 and a contractile prostanoid antagonist, L-640,305. These results demonstrate that F-met-leu-phe is potent bronchoconstrictor in the guinea pig and that the peptide may induce these changes through the generation of products of the cyclooxygenase and lipoxygenase pathways of arachidonic acid metabolism.