bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Th2 responses in the development of allergen‐induced airway remodelling in a murine model of allergic asthma
Authors: Komai Masato , Tanaka Hiroyuki , Masuda Taisei , Nagao Koichi , Ishizaki Masayuki , Sawada Masatsugu , Nagai Hiroichi 
Published: March 2003

To clarify the involvement of Th2 responses in the development of allergen‐induced airway remodelling, we investigated the effect of anti‐CD4 monoclonal antibody (mAb) and anti‐CD8 mAb, and the responses of IL‐4 gene‐knockout (KO) mice in a murine model of allergic asthma.
Mice were immunized twice by intraperitoneal injections of ovalbumin (OA), and exposed to aeroallergen (OA, 1% w v−1) for 3 weeks. Twenty‐four hours after the final challenge, airway responsiveness to acetylcholine was measured, and bronchoalveolar lavage (BAL) and histological examinations were carried out.
Anti‐CD4 mAb (1 mg kg−1) clearly inhibited allergen‐induced increases in airway responsiveness to acetylcholine, the number of eosinophils in BAL fluid, serum OA‐specific IgE levels, IL‐13 and transforming growth factor‐β1 levels in BAL fluid, and amount of hydroxyproline in the lung by 100, 99, 100, 100, 84, and 60%, respectively. Furthermore, the antibody (1 mg kg−1) also attenuated allergen‐induced goblet cell hyperplasia in the epithelium and subepithelial fibrosis by 72 and 83%, respectively. In contrast, anti‐CD8 mAb (1 mg kg−1) showed no effect on each parameter. Furthermore, all these parameters were attenuated in IL‐4KO mice by 57, 93, 100, 45, 84 and 60%, and also 72 and 83%, respectively.
These findings suggest that Th2 responses play a critical role for the development of allergen‐induced airway remodelling, and that the inhibition of Th2 responses, e.g. using anti‐CD4 mAb, is a therapeutic approach for the treatment of airway remodelling in asthma.

British Journal of Pharmacology (2003) 138, 912–920. doi:10.1038/sj.bjp.0705105
Protease‐activated receptor (PAR)‐independent growth and pro‐inflammatory actions of thrombin on human cultured airway smooth muscle
Authors: Tran Thai , Stewart Alastair G 
Published: March 2003

Thrombin, a mitogen for human cultured airway smooth muscle (HASM), has many actions that have been attributed to activation of protease‐activated receptor (PARs). However, the role of PARs in the proliferative action has not been clearly identified. Moreover, thrombin elicits cytokine production in a number of cell types, but these effects have not been characterized in human ASM.
Thrombin (0.03–3 U ml−1)‐stimulated increases in the levels of the pro‐inflammatory and fibrogenic cytokine, granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) were observed over the same concentration range observed for thrombin‐stimulated mitogenesis.
Inhibition of thrombin proteolytic activity, with either D‐phenylalanyl‐L‐prolyl‐L‐arginine chloromethyl ketone (PPACK)‐ or hirudin‐treated thrombin (0.3 U ml−1) or in the presence of the thrombin serine protease‐selective inhibitor, SDZ 217‐766 (0.15 μM), reduced the thrombin‐stimulated GM‐CSF levels by 91±3, 65±12 and 83±9% (n=8, P<0.05), respectively. PPACK treatment, hirudin and SDZ 217‐766 inhibited thrombin‐stimulated increase in cell number by 70±8, 63±11 and 69±8%, respectively.
PAR‐selective peptides SFLLRN (PAR1; 10 μM), SLIGKV (PAR2; 10 μM), GYPGQV (PAR4; 100 μM) or the combination of SFLLRN and GYPGQV elicited mitogenic responses of only 15% of that to thrombin and surprisingly, had no effect on GM‐CSF levels (n=8). Nevertheless, inhibition of thrombin responses by pertussis toxin (50 ng ml−1) suggests that the PAR‐independent actions also involve a G‐protein‐coupled receptor.
PAR1 receptor expression was evident by immunohistochemistry and these receptors were coupled to increases in intracellular calcium, but not to the phosphorylation of ERK or the increases in cyclin D1 protein levels that are essential for cell proliferation. Cross‐desensitization of intracellular calcium increases by thrombin and the PAR1‐selective peptide provides evidence that the PAR1 receptor responds to both ligands.
The failure of PAR‐selective peptides to mimic thrombin responses together with the inhibition of thrombin responses by serine protease inhibitors suggest the involvement of novel proteolytic receptor targets for thrombin‐induced mitogenesis and cytokine production.

British Journal of Pharmacology (2003) 138, 865–875. doi:10.1038/sj.bjp.0705106
Local anaesthetics have different mechanisms and sites of action at the recombinant N‐methyl‐D‐aspartate (NMDA) receptors
Authors: Sugimoto Masahiro , Uchida Ichiro , Mashimo Takashi 
Published: March 2003

Although the principal pharmacological targets of local anaesthetics (LAs) are voltage‐gated Na+ channels, other targets have also been suggested. Here we examined the effects of LAs on the N‐methyl‐D‐aspartate (NMDA) receptor, a receptor involved in the process of nociception.
LAs (bupivacaine, lidocaine, procaine, and tetracaine) reversibly and concentration‐dependently inhibited recombinant ε1/ζ1 and ε2/ζ1 NMDA receptors expressed in Xenopus oocytes (IC50s for bupivacaine, lidocaine, procaine, and tetracaine were 1032.0, 1174.1, 642.1 and 653.8 μM at the ε1/ζ1 receptor; and 1090.8, 1821.3, 683.0 and 662.5 μM respectively (at the ε2/ζ1 receptor). Bupivacaine and procaine were non‐competitive antagonists; bupivacaine possesses non‐competitive and competitive actions when interacting with glycine, whereas procaine has only non‐competitive action.
Mutation of asparagine residue at position 598 (Asp598) in the ζ1 subunit, a residue associated with the blockade site for Mg2+ and ketamine, to glutamine or arginine reduced the sensitivity to procaine but not to bupivacaine. Thus, procaine may interact with sites of action that are closely related to those of Mg2+ and ketamine blockade.
These results suggest that LAs inhibit the NMDA receptor by various mechanisms.

British Journal of Pharmacology (2003) 138, 876–882. doi:10.1038/sj.bjp.0705107
Role of nuclear factor‐κB activation in acute ischaemia‐reperfusion injury in myocardium
Authors: Kis Adrienn , Yellon Derek M , Baxter Gary F 
Published: March 2003

Our aims were to characterize activation of the transcription factor, nuclear factor kappa‐B(NF‐κB), during myocardial ischaemia‐reperfusion and to assess its functional role in the evolution of acute ischaemia‐reperfusion injury in intact myocardium in vivo.
Under pentobarbitone anaesthesia, rabbits underwent sham operation, 30 min left coronary artery occlusion followed by 0, 10 or 180 min reperfusion. Saline or NF‐κB inhibitor diethyldithiocarbamic acid (DDTC, 50, 100 or 200 mg kg−1) was given intravenously 5 min prior to reperfusion.
Electromobility shift assay revealed that 30 min ischaemia alone did not activate NF‐κB compared to time‐matched sham‐operated controls (85±13% vs 100±28%, respectively). However, ischaemia plus 10 min reperfusion markedly increased activation of NF‐κB (295±77%). DDTC 50 mg kg−1 did not inhibit NF‐κB activation (278±67%) but at the higher doses complete inhibition was observed (54±20%, 31±16%, respectively).
Infarct to risk ratio was determined by triphenyltetrazolium chloride staining after 30 min ischaemia and 180 min reperfusion. DDTC 50 or 100 mg kg−1 significantly reduced infarct size compared to the saline‐treated control group (34.9±5.2%, 37.1±5.9%, vs 51.3±3.6%, P<0.05, respectively), whereas there was no protection with 200 mg kg−1 (45.6±5.3%).
We conclude that ischaemia alone does not activate NF‐κB, but post‐ischaemic reperfusion robustly activates NF‐κB in the myocardium. DDTC limited irreversible injury at low doses, but this effect appears to be dissociated from inhibition of NF‐κB. Thus, activation of NF‐κB during reperfusion does not appear to play a role in the evolution of myocardial infarction during the early phase of reperfusion.

British Journal of Pharmacology (2003) 138, 894–900. doi:10.1038/sj.bjp.0705108
Expression and functional role of Rho‐kinase in rat urinary bladder smooth muscle
Authors: Wibberley Alexandra , Chen Zunxuan , Hu Erding , Hieble J Paul , Westfall Timothy D 
Published: March 2003

The involvement of Rho‐kinase (ROCK) in the contractile mechanisms mediating smooth muscle contraction of the rat urinary bladder was investigated using expression studies and the ROCK inhibitor Y‐27632.
Both isoforms of ROCK (ROCK I and ROCK II) were detected in high levels in rat urinary bladder.
Y‐27632 (10 μM) significantly attenuated contractions of rat urinary bladder strips evoked by the G‐protein coupled receptor agonists carbachol (58.1±10.5% at 0.3 μM) and neurokinin A (68.6±12.7% at 1 μM) without affecting contractions to potassium chloride (10–100 mM). In addition, basal tone was reduced by 47.8±2.0% by 10 μM Y‐27632 in the absence of stimulation.
Contractions of urinary bladder strips evoked by the P2X receptor agonist α,β‐methylene ATP (α,β‐mATP; 10 μM) were also attenuated by Y‐27632 (30.0±7.2% at 10 μM).
Y‐27632 (10 μM) significantly attenuated contractions evoked by electrical field stimulation (2–16 Hz). The effect of Y‐27632 on the tonic portion of the neurogenic response (4–16 Hz) was not significantly different from the effect of atropine (1 μM) alone.
While the mechanism underlying the ability of Y‐27632 to inhibit α,β‐mATP‐evoked contractions remains undetermined, the results of the present study clearly demonstrate a role for ROCK in the regulation of rat urinary bladder smooth muscle contraction and tone.

British Journal of Pharmacology (2003) 138, 757–766. doi:10.1038/sj.bjp.0705109
Neurogenic responses mediated by vanilloid receptor‐1 (TRPV1) are blocked by the high affinity antagonist, iodo‐resiniferatoxin
Authors: Rigoni Michela , Trevisani Marcello , Gazzieri David , Nadaletto Riccardo , Tognetto Michele , Creminon Christophe , Davis John B , Campi Barbara , Amadesi Silvia , Geppetti Pierangelo , Harrison Selena 
Published: March 2003

Stimulation of the vanilloid receptor‐1 (TRPV1) results in the activation of nociceptive and neurogenic inflammatory responses. Poor specificity and potency of TRPV1 antagonists has, however, limited the clarification of the physiological role of TRPV1.
Recently, iodo‐resiniferatoxin (I‐RTX) has been reported to bind as a high affinity antagonist at the native and heterologously expressed rat TRPV1. Here we have studied the ability of I‐RTX to block a series of TRPV1 mediated nociceptive and neurogenic inflammatory responses in different species (including transfected human TRPV1).
We have demonstrated that I‐RTX inhibited capsaicin‐induced mobilization of intracellular Ca2+ in rat trigeminal neurons (IC50 0.87 nM) and in HEK293 cells transfected with the human TRPV1 (IC50 0.071 nM).
Furthermore, I‐RTX significantly inhibited both capsaicin‐induced CGRP release from slices of rat dorsal spinal cord (IC50 0.27 nM) and contraction of isolated guinea‐pig and rat urinary bladder (pKB of 10.68 and 9.63, respectively), whilst I‐RTX failed to alter the response to high KCl or SP.
Finally, in vivo I‐RTX significantly inhibited acetic acid‐induced writhing in mice (ED50 0.42 μmol kg−1) and plasma extravasation in mouse urinary bladder (ED50 0.41 μmol kg−1).
In in vitro and in vivo TRPV1 activated responses I‐RTX was ∼3 log units and ∼20 times more potent than capsazepine, respectively. This high affinity antagonist, I‐RTX, may be an important tool for future studies in pain and neurogenic inflammatory models.

British Journal of Pharmacology (2003) 138, 977–985. doi:10.1038/sj.bjp.0705110
Apoptosis and necrosis: two distinct events induced by cadmium in cortical neurons in culture
Authors: López E , Figueroa S , Oset‐Gasque M J , González M P 
Published: March 2003

Cadmium is an extremely toxic metal commonly found in industrial workplaces, a food contaminant and a major component of cigarette smoke. Cadmium can severely damage several organs, including the brain. In this work, we have studied both the cadmium toxicity on rat cortical neurons in culture and the possible protective effect of serum.
Our results indicate that: (1) cadmium is taken up by the neurons in a dose and serum dependent way; (2) cadmium, at concentrations from 1 μM or 10 μM (depending on the absence or the presence of serum) up to 100 μM, decreases the metabolic capacity, which was evaluated by the XTT (tetrazolium salt) test; (3) cadmium induces apoptosis and LDH (lactate dehydrogenase) release in a dose dependent way; (4) in a serum‐free medium, the cadmium‐induced apoptosis is accompanied by caspase‐3 activation; (5) both the caspase‐3 activation and the cadmium‐induced apoptosis are reversed by N‐acethyl‐Asp‐Glu‐Val‐Asp‐aldehyde (Ac‐DEVD‐CHO), a selective caspase‐3 inhibitor, indicating that the caspase‐3 pathway is involved in cadmium‐induced apoptosis in cortical neurons; and (6) the cadmium concentrations which produce caspase‐3 activation do not modify the intracellular ATP levels; however, higher cadmium concentrations lead to both intracellular ATP depletion and ATP release, but do not increase the caspase‐3 activity, indicating that cadmium also produces cellular death by necrosis.
These results suggest that cadmium induces either apoptosis or necrosis in rat cortical neurons, depending on the cadmium concentration.

British Journal of Pharmacology (2003) 138, 901–911. doi:10.1038/sj.bjp.0705111
Inhibition of cyclic GMP hydrolysis with zaprinast reduces basal and cyclic AMP‐elevated L‐type calcium current in guinea‐pig ventricular myocytes
Authors: Ziolo Mark T , Lewandowski Susanne J , Smith Jacquelyn M , Romano Fred D , Wahler Gordon M 
Published: March 2003

Cyclic GMP (cGMP) has been shown to be an important modulator of cardiac contractile function. A major component of cGMP regulation of contractility is cGMP‐mediated inhibition of the cardiac calcium current (ICa). An under‐appreciated aspect of cyclic nucleotide signalling is hydrolysis of the cyclic nucleotide (i.e., breakdown by phosphodiesterases (PDEs)). The role of cGMP hydrolysis in regulating ICa has not been studied. Thus the purpose of this study was to investigate if inhibition of cGMP hydrolysis can modulate ICa in isolated guinea‐pig ventricular myocytes.
Zaprinast, a selective inhibitor of cGMP‐specific PDE (PDE5), caused a significant increase in cGMP levels in myocytes, but was without affect on basal or β‐adrenergic stimulated cAMP levels (consistent with its actions as a specific inhibitor of PDE5).
Zaprinast inhibited ICa that was pre‐stimulated with cAMP elevating agents (isoproterenol, a β‐adrenergic agonist; or forskolin, a direct activator of adenylate cyclase). The effect of zaprinast was greatly reduced by KT5823, an inhibitor of cGMP‐dependent protein kinase (PKG).
Zaprinast also significantly inhibited basal ICa when perforated‐patch or whole‐cell recording with physiological pipette calcium concentration (10−7 M) was used. However, this effect was not observed when using standard calcium‐free whole‐cell recording conditions.
These results indicate that inhibition of cGMP hydrolysis can decrease both basal and cAMP‐stimulated ICa. Thus, cGMP hydrolysis may likely be an important step for physiological modulation of ICa. This regulation may also be important in disease states in which cGMP production is increased and PDE5 expression is altered, such as heart failure.

British Journal of Pharmacology (2003) 138, 986–994. doi:10.1038/sj.bjp.0705112
RXR activators molecular signalling: involvement of a PPARα‐dependent pathway in the liver and kidney, evidence for an alternative pathway in the heart
Authors: Ouamrane Laïla , Larrieu Gilberte , Gauthier Béatrice , Pineau Thierry 
Published: March 2003

In this study we compared the molecular signalling elicited by rexinoids, selective retinoid X receptor (RXR)‐activators, in several organs (i.e. liver, kidney, heart) and in hepatocytes of various species.
RXR plays the pivotal role of a hetero‐dimerization partner for the members of the class II subset of nuclear receptors which regulate the transcription of numerous target genes, following chemical activation. Several of these selective activators are currently used to treat hyperlipidaemia (fibrates), type II diabetes (glitazones), or skin disorders (retinoic acid). Although these therapeutic pathways are not fully elucidated, receptor activation is considered a pre‐requisite for efficacy. Therefore RXR, which accepts numerous dimeric partners, is considered a worthwhile pharmacological target.
We analysed a number of biochemical and molecular responses to rexinoids which were given orally to mice. Our results showed a prominent involvement of the peroxisome proliferator‐activated receptor (PPARα) as a majority of the observed hepatic and renal regulations were abolished in PPARα‐knockout animals. Therefore we documented the species‐specificity of these rexinoid actions which were reproduced in rat primary hepatocyte cultures but not in cultures of rabbit or human origin. Conversely, we established that the regulation of the pyruvate dehydrogenase kinase (PDK4) gene in the heart, by rexinoids, is independent of PPARα expression.
Our results support the obligatory expression of the active, although quiescent, PPARα to sustain a subset of relevant regulations attributable to rexinoids in the liver and kidney. Their cardiac molecular signalling unveiled an alternate transduction pathway and therefore opens new prospects in the therapeutic potential of rexinoids.

British Journal of Pharmacology (2003) 138, 845–854. doi:10.1038/sj.bjp.0705113
Cannabinoid receptor‐independent inhibition by cannabinoid agonists of the peripheral 5‐HT3 receptor‐mediated von Bezold–Jarisch reflex
Authors: Godlewski Grzegorz , Göthert Manfred , Malinowska Barbara 
Published: March 2003

On the basis of previous findings that cannabinoids inhibit the function of human and rat 5‐HT3 receptors in vitro, we investigated whether cannabinoid receptor agonists also modulate the activity of the rat peripheral 5‐HT3 receptors on the terminals of cardiopulmonary afferent C‐fibres in vivo.
In urethane‐anaesthetized rats, pre‐treated intravenously (i.v.) with the CB1 receptor antagonist SR 141716A (3 μmol kg−1) and with the β1/β2 adrenoceptor antagonist propranolol (0.3–0.4 μmol kg−1), bolus injection of the serotonin 5‐HT3 receptor agonist phenylbiguanide (3–10 μg kg−1, i.v.) or the vanilloid VR1 receptor agonist capsaicin (3–10 μg kg−1, i.v.) caused an immediate decrease in heart rate and mean arterial blood pressure (the von Bezold–Jarisch reflex).
The phenylbiguanide‐induced bradycardia was dose‐dependently attenuated by the cannabinoid receptor agonists CP 55,940 (0.1–1 μmol kg−1, i.v.) and WIN 55,212‐2 (0.1–3 μmol kg−1, i.v.) 20 min after injection, but not by the inactive S‐(−)enantiomer of the latter, WIN 55,212‐3 (1 μmol kg−1, i.v.). The inhibition was reversible within 30 min. The extent of inhibition by the highest doses of cannabinoid receptor agonists amounted to about 50%. Both cannabinoid receptor agonists failed to affect the capsaicin‐evoked bradycardia.
In conclusion, our results demonstrate that cannabinoid receptor agonists modulate the von Bezold–Jarisch reflex by inhibiting peripheral serotonin 5‐HT3 receptors in rats in vivo. An analogous mechanism of cannabinoid receptor agonists may be assumed to be involved in other serotonin 5‐HT3 receptor‐mediated responses.

British Journal of Pharmacology (2003) 138, 767–774. doi:10.1038/sj.bjp.0705114
A photosensitive vascular smooth muscle store of nitric oxide in mouse aorta: no dependence on expression of endothelial nitric oxide synthase
Authors: Andrews Karen L , McGuire John J , Triggle Chris R 
Published: March 2003

Photorelaxation is the reversible relaxation of vascular smooth muscle (VSM) when irradiated with ultraviolet (UV) light resulting from the release of nitric oxide (NO). In this study we characterize the involvement of endothelial nitric oxide synthase (eNOS) in the photorelaxation response of thoracic aorta from endothelial NOS deficient (−/−) and control (C57BL/6j) mice.
Cirazoline contracted aortae were repeatedly exposed to 30 s of UV light every 3–4 min. Equal levels of photorelaxation (45±2%; n=34) was observed in both strains.
1H‐[1,2,4]‐oxadiazolo[4,3‐a]quinoxalin‐1‐one (ODQ), K+, 2‐(4‐carboxyphenyl)‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl‐3‐oxide (c‐PTIO), 4‐aminopyridine (4‐AP) and ethacrynic acid significantly reduced the photorelaxation response. In C57BL/6j mice diethyldithiocarbamate (DETCA) also reduced photorelaxation.
Control endothelium‐intact and ‐denuded aorta and L‐NAME (100 μM) treated and untreated eNOS (−/−) aortae were repeatedly exposed to UV light for 5 min every 10 min until no photorelaxation response was observed. After 1 h of rest in the dark the vessels showed between 30–70% recovery of the photorelaxation response indicating regeneration of the store in the absence of the endothelium and eNOS.
The results of this study suggest that photorelaxation in mouse aorta VSM results from the release of NO from a stable store of RSNOs, which activates soluble guanylate cyclase (sGC), leading to cGMP‐dependent relaxation that is partially mediated by an increase in KV channel activation and hyperpolarization. In addition, the eNOS isoform is not essential for the formation of the photorelaxation store and a non‐NOS source of NO may be involved in the maintenance of this store.

British Journal of Pharmacology (2003) 138, 932–940. doi:10.1038/sj.bjp.0705115
Functional coupling of the human dopamine D2 receptor with Gαi1, Gαi2, Gαi3 and Gαo G proteins: evidence for agonist regulation of G protein selectivity
Authors: Gazi Lucien , Nickolls Sarah A , Strange Philip G 
Published: March 2003

The human dopamine D2long (D2L) receptor was expressed with four different G proteins in Sf9 cells using the baculovirus expression system. When co‐expressed with Gi/Go G proteins (Gi1α, Gi2α, Gi3α, or Goα, plus Gβ1 and Gγ2), the receptor displayed a high‐affinity binding site for the agonists (dopamine and NPA), which was sensitive to GTP (100 μM), demonstrating interaction between the receptor and the different G proteins.
The receptor to G protein ratio (R : G ratio) was evaluated using [3H]‐spiperone saturation binding (R) and [35S]‐GTPγS saturation binding (G). R : G ratios of 1 : 12, 1 : 3, 1 : 14 and 1 : 5 were found for Gi1, Gi2, Gi3, and Go preparations, respectively. However, when R : G ratios of 1 : 2 and 1 : 12 were compared for Gi2 and Go, no difference was found for the stimulation of [35S]‐GTPγS binding.
Several agonists were tested for their ability to stimulate [35S]‐GTPγS binding to membranes co‐expressing the receptor and various G proteins. All the compounds tested showed agonist activity in preparations expressing Gi3 and Go. However, for Gi2 and Gi1 preparations, compounds such as S‐(−)‐3‐PPP and p‐tyramine were unable to stimulate [35S]‐GTPγS binding.
Most of the compounds showed higher relative efficacies (compared to dopamine) and higher potencies in the preparation expressing Go. Comparison of the effects of different agonists in the different preparations showed that each agonist differentially activates the four G proteins.
We conclude that the degree of selectivity of G protein activation by the D2L receptor can depend on the conformation of the receptor stabilised by an agonist.

British Journal of Pharmacology (2003) 138, 775–786. doi:10.1038/sj.bjp.0705116