European Journal of Pharmacology: Environmental Toxicology and Pharmacology最新文献

This study was undertaken to compare the tumour promoting effects induced by 3,4,5,3′,4′-pentachlorobiphenyl (PCB 126) and 2,3,7,8,-tetrachlorodibenzo-p-dioxin (TCDD). In addition, interactive effects in rats treated with combinations of PCB 126 and TCDD were studied. Partially hepatectomized female Sprague-Dawley rats were initiated with nitrosodiethylamin. After 5 weeks of recovery the promotion treatment started and continued for 20 weeks. The results from the present study demonstrate that PCB 126 elicit approximately 10% of TCDD's tumour promoting activity measured as enhancement of the development of γ-glutamyl-transpeptidase-positive altered heaptic foci in the liver. The factor required for the PCB to match the response of TCDD was adopted as a toxic equivalency factor and was in this case 0.1, which is the same as the factor suggested by Ahlborg et al. (1994).In the groups treated with a mixture of PCB 126 and TCDD the tumour promoting effect indicated an additive response. This result suggests that PCB 126 and TCDD act by the same mechanistical pathway, which in turn, supports that the toxic equivalency factor-concept can be used for TCDD-like tumour promoters.

To investigate cellular electrophysiologic alterations due to lipid peroxidation of the cell membrane by free radicals as a possible cause of coronary reperfusion arrhythmias, we studied the effects of t-butyl hydroperoxide on the spontaneous action potential and membrane currents of the rabbit sinoatrial and atrioventricular node preparations (0.2 × 0.2 × 0.1 mm). 1–5 min of superfusion with t-butyl hydroperoxide (100–500 μM) caused a transient increase in the spontaneous firing frequency by 9%, accompanied by a 4% increase in the action potential amplitude and a 33% increase in the maximal rate of depolarization (P<0.05, n = 6). t-Butyl hydroperoxide then gradually suppressed physiological automaticity, but induced abnormal repetitive firing due to early and delayed after-depolarizations. 15 min of superfusion with t-butyl hydroperoxide caused a complete standstill of nodal cells at a resting potential of −46 ± 3 mV (n = 12). Such effects of t-butyl hydroperoxide on the spontaneous action potential were attenuated by pretreating the cells with butylated hydroxytoluene, a lipid peroxidation inhibitor. Voltage clamp experiments using double microelectrode methods revealed that t-butyl hydroperoxide transiently increased the Ca²⁺ current by 22% after 5 min of superfusion but subsequently reduced it to 46% of the control value after 15 min (P<0.05, n = 6). Similar biphasic changes were observed in the delayed rectifying K⁺ current and hyperpolarization-activated inward current (n = 6). Background current was progressively increased without any change in its reversal potential (n = 6). These results suggest that membrane lipid peroxidation may accelerate or suppress physiological automaticity and induce abnormal automaticity by both impairing cellula metabolic function and damaging the lipid membrane structure as well as ionic channel protein.

The time course of the effects of ethanol alone and in combination with the selective α₂-adrenoceptor agonist dexmedetomidine and the α-adrenoceptor antagonist atipamezole was studied in NIH-Swiss mice. Core body temperature, rotarod performance, motility and changes in the noradrenaline, dopamine, and 5-hydroxytryptamine (5-HT) metabolite contents of different brain parts (limbic forebrain, striatum, lower brainstem, the rest of the forebrain + midbrain and hypothalamus) were measured. Atipamezole (3 mg/kg) attenuated the hypothermia induced by either ethanol (3 g/kg) alone or ethanol in combination with dexmedetomidine (0.3 mg/kg). Atipamezole shortened the duration of the ethanol-impaired and ethanol + dexmedetomidine-impaired rotarod performance. Further, atipamezole prevented the decreased motility due to the combined treatment with ethanol and dexmedetomidine. Ethanol increased 3-methoxy-4-hydroxyphenylethylene glycol (MHPG), homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) values. Dexmedetomidine alone decreased MHPG and 5-hydroxyindoleacetic acid (5-HIAA) concentrations and increased DOPAC and HVA values. Dexmedetomidine combined with ethanol resulted in a further increase in DOPAC and HVA values. Pharmacokinetic parameters did not contribute to this antagonism of ethanol's effects by atipamezole, nor did the antagonism observed in rotarod performance or hypothermia seem to correlate with the changes seen in the brain noradrenaline and dopamine or 5-HT metabolism. In conclusion, these findings suggest that several ethanol effects are not mediated via direct activation of α₂-adrenoceptors, even though some of ethanol's behavioral and physiological effects may be antagonized by coadministration of α₂-adrenoceptor antagonists.

The protein glycosylation inhibitor tunicamycin protected male BALB/c mice from tumor necrosis factor α-induced liver failure. Tunicamycin also inhibited tumor necrosis factor-induced cell death in primary hepatocyte cultures with a median inhibitory concentration of 8 nM, but not in the tumor cell line WEHI 164 clone 13. Hepatocyte death in our culture system was characterized by DNA fragmentation and apoptotic changes. These two characteristic signs of programmed cell death were also inhibited by tunicamycin treatment. These data suggest that protein glycosylation is an early and causal event of tumor necrosis factor (TNF)-induced parenchymal cell death in the liver.

We have confirmed our earlier finding that most red wines are able to bring about 5-hydroxytryptamine (5-HT, serotonin) release from platelets in vitro. Platelets from individual subjects manifested varying degrees of releasing ability but responded to different wines with a similar rank ordering. There was a high correlation (r = 0.87) between the effect of red wine and that of reserpine in different individuals. Some types of red wine caused a consistently higher release of 5-HT than others in all subjects; one red wine in particular resulted in neglible release. When several brands of this ·low-releasing” red wine were further examined, they all showed a lower activity than all the brands of a ‘high-releasing’ red wine type. This variation in releasing power was not related to intensity of red colour. Partial purification of red wine was achieved by column chromatography and showed releasing activity to be associated with a low molecular weight orange fraction. Preliminary studies, using solid phase extraction methods, showed that the active components lie mainly in a subgroup of the flavonoid fraction. If any of the adverse effects of red wine, such as headache induction, derive from this 5-HT releasing ability, then it may be possible to prepare red wines free from the chemical substances responsible.

At which time-point and to what extent do $\text{N-}\text{methyl}\text{-}\text{d}\text{-}\text{aspartate}$ (NMDA) receptors, α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/kainate receptors and L-type voltage-sensitive Ca²⁺ channels (VSCC) contribute to glutamate-induced neuronal injury? To address this question, we induced glutamate neurotoxicity in two neuronal culture systems, chick telencephalic neurons and rat hippocampal neurons, and tested selective antagonists for their neuroprotective activity when administered either during the excitotoxic insult (acute treatment) or during the recovery period (posttreatment). In cultured chick telencephalic neurons exposed to 1 mM l-glutamate for 60 min, both the NMDA receptor antagonist dizocilpine (MK-801; 0.1 μM) and the AMPA/kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 1 μM) completely blocked glutamate-induced neuronal injury when applied concomitantly with glutamate. If the antagonists were applied during the recovery period, dizocilpine at concentrations up to 10 μM only moderately increased cell viability, whereas CNQX showed a neuroprotective activity comparable to that observed in the case of the acute treatment. In cultured rat hippocampal neurons, excitotoxic injury was induced by a 30-min exposure to 1 mM glutamate. Treatment with dizocilpine during the glutamate exposure could rescue the hippocampal neurons from the excitotoxic insult, whereas acute treatment with the AMPA/kainate receptor antagonist 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(F)-quinoxalime (NBQX) or the L-type VSCC blocker nimodipine showed no protection. In contrast, all three drugs showed neuroprotective activity when applied 30, 60 or 120 min after the glutamate exposure. Surprisingly, when the onset of the treatment was delayed for even 240 min, only NBQX and nimodipine led to a reduction in excitotoxic neuronal injury. We conclude that activation of AMPA/kainate receptors and L-type VSCC is critically involved in a late stage of glutamate neurotoxicity, thereby allowing pharmacological intervention at a time when blockade of NMDA receptors becomes less efficacious.

A human cell subline (PC-9 / VCR) resistant to vincristine was established from non-small cell lung cancer PC-9 cells by incremental exposure of the cells to vincristine. The resistant cells showed phenotypic resistance to vincristine (10-fold), colchicine (6.9-fold) and cisplatin (1.4-fold) but they showed sensitivity to other chemotherapeutic agents including melphalan and etoposide VP-16. The characteristics of the vincristine resistance was partially inhibited (5–7-fold) by co-treatment of PC-9/VCR cells with a nontoxic concentration of L-ascorbic acid (25 μg/ml). Co-treatment or 96 h pre-treatment with ascorbic acid resulted in potentiation of the vincristine effect on the resistant, but not on the sensitive, cell line. The growth inhibition due to vincristine treatment after 24 or 96 h growth in ascorbic acid-free medium was decreased in the resistant as well as in the sensitive cell line. In both cell lines, enhanced growth rate has been shown after ascorbic acid treatment. Similarly, cross-resistance of PC-9/VCR cells to colcholine also be blocked by ascorbic acid. In addition, a nontoxic concentration of verapamil, a known multidrug resistance inhibitor, did not affect the resistant phenotype of PC-9/VCR cells. These findings suggest that an ascorbic acid-sensitive mechanism may be involved in drug resistance per se in the human lung cancer cells, which differs from the classical phosphoglycoprotein-mediated or previously reported non-phosphoglycoprotein-mediated multidrug resistance.

The effects of ethanol on L-type Ca²⁺ and fast Na⁺ currents (I_Ca, and I_Na, respectively) were examined using the whole-cell patch-clamp experiments on guinea-pig ventricular cells. At a clinically relevant concentration of 24 mM, ethanol slightly but significantly shortened the action potential duration, and reduced the I_Ca by 7 ± 4% (mean ± S.D.). This concentration of ethanol did not affect I_Na, but a lethal concentration of ethanol (80 mM) significantly inhibited I_Na by 13 ± 5%. The voltage dependence of I_Na activation was not affected by ethanol, whereas the inhibitions of I_Ca by 80 mM ethanol and I_Na by 240 mM were both accompanied by a several mV shift in the channel availability curve toward more negative potentials, suggesting that the channels in the inactivated state are more susceptible to ethanol. The I_Ca inhibition by ethanol at clinically relevant concentrations could contribute to a negative inotropic effect, action potential shortening and development of arrhythmias, while the pathophysiological significance of ethanol inhibition of I_Na seems less important.

β-Eudesmol, a sesquiterpenol present in Chinese herbs, improved the tetanic contraction impaired by diisopropylfluorophosphate in isolated mouse diaphragm preparations by an inhibition of the regenerative acetylcholine release. The antagonism was enhanced when a small concentration of obidoxime was present. Neither enzyme reactivation nor curare-like action was evident. β-Eudesmol (300 mg/kg, i.p.) elevated the LD₅₀ of diisopropylfluorophosphate (s.c.) in control mice from 4.2 to 6.4 mg/kg and in mice pretreated with atropine from 7.8 to 10.6 mg/kg. In mice pretreated with atropine and obidoxime, β-eudesmol showed a greater synergistic effect, increasing the LD₅₀ from 281 to more than 800 mg/kg. β-Eudesmol also markedly alleviated diisopropylfluorophosphate-induced muscle fasciculation, tremor and convulsion and prolonged the time to death. It is proposed that β-eudesmol may be added to the standard antidotal regimen (atropine plus obidoxime) for treating organophosphate intoxication.

Trimellitic anhydride is a cause of occupational asthma in humans. We have previously found that tracheal instillation of trimellitic anhydride conjugated to guinea pig serum albumin induces acute bronchoconstriction and airway plasma exudation in sensitised animals, responses mediated primarily via histamine release. In the present study, neural mechanisms mediating bronchoconstriction and goblet cell secretion were determined in trimellitic anhydride-sensitised guinea pigs using the ganglionic blocker hexamethonium to eliminate efferent reflex mechanisms, pretreatment with capsaicin to eliminate afferent mechanisms, or cimetidine and mepyramine to eliminate histamine-mediated mechanisms. The magnitude of secretion of intracellular mucus from tracheal goblet cells was quantified morphometrically as a mucus score which is inversely related to the degree of discharge. Guinea pigs were injected intradermally either with 0.1 ml 0.3% trimellitic anhydride in corn oil or with corn oil alone as control. Fourteen to eighteen days later all sensitised animals had developed specific immunoglobulin (Ig) G1 antibodies whereas the controls had not. Tracheal instillation of conjugated trimellitic anhydride in anaesthitised animals significantly increased airway lung resistance (R_L) 24-fold in sensitised guinea pigs (34.3 ± 7.9 cm H₂O · ml⁻¹ · s) compared with controls (1.4 ± 0.1 cm H₂O · ml⁻¹ · s). Mucus score was significantly reduced by 51% (indicating goblet cell secretion) in sensitised guinea pigs (183 ± 22 mucus score units) compared with controls (372 ± 41 mucus score units). The antihistamines significantly inhibited conjugated trimellitic anhydride-induced bronchoconstriction by 89%, but did not significantly affect goblet cell discharge. Hexamethonium alone did not significantly affect conjugated trimellitic anhydride-induced bronchoconstriction or goblet cell secretion. Capsaicin pretreatment (in combination with hexamethonium) significantly inhibited golet cell discharge (by 80%) but had no significant effect on bronchoconstriction. We conclude that conjugated trimellitic anhydride challenge of trimellitic anhydride-sensitised guinea pigs induces goblet cell discharge and bronchoconstriction via different mechanisms with activation of capsaicin-sensitive sensory nerves responsible for secretion and histamine release responsible for airway constriction. The guinea pig model of trimellitic anhydride-induced occupational asthma may prove useful in examination of mechanisms of goblet cell secretion in allergic diseases.