The action of caffeine on inward barium current through voltage-dependent calcium channels in single rabbit ear artery cells

The effect of caffeine on inward current carried by barium ions through voltage-dependent calcium channels has been investigated in single rabbit ear artery cells using whole-cell voltage-clamp techniques. Caffeine (1 –30 mM) caused a rapid and reversible concentration-dependent blockade of barium current and a related compound, 3-isobutyl-1-methylxanthine (IBMX), was a more potent inhibitor of barium current. Caffeine-induced inhibition of barium current showed no voltage- or usedependence and caffeine did not alter the steady-state inactivation of barium current. The effect of caffeine was not blocked by extracellular or by intracellular ryanodine or inclusion of both 5 mM 1,2-bis(2-aminophenoxy)-ethane N,N,N,N,-tetraacetic acid (BAPTA) and 2 mM ethylene glycol-bis(-amino ethyl ether) N,N,N,N,-tetraacetic acid (EGTA) in the intracellular solution. Rolipram and M&B 22984, non-xanthine inhibitors of phosphodiesterase, did not diminish inward barium current. The data indicate that caffeine and IBMX block voltage-operated calcium channels and it is suggested that this is due to a direct interaction of methylxanthines with the calcium channel.     

Independent interaction of alprazolam and caffeine under chronic dose regimens on differential reinforcement of low-rate (DRL 45-s) performance

In previous research, we found an independent interaction of alprazolam and caffeine in rats under acute dose regimens using two measures (reinforcement rate and shorter-response rate) of a differential reinforcement of low rate performance (DRL 45-s) in 3-h sessions. Applying the same behavioral endpoints, the present study investigated the alprazolam-caffeine interaction under chronic dose regimens. Both drugs were administered by the oral route. Acute alprazolam and caffeine dose-response curves (DRCs) were characterized and were then used to determine the maintenance dose for the respective chronic dose regimens. Both drugs decreased the reinforcement rate and increased the shorter-response rate in a dose-related fashion. An alprazolam DRC also was determined during chronic-caffeine, chronic-alprazolam, and concurrent chronic-caffeine-alprazolam dose regimens. Complete tolerance to caffeine-induced rate changes was observed on the second day. Incomplete tolerance occurred only at higher alprazolam doses (7–12.5 mg/kg). Cross tolerance was not found between alprazolam and caffeine. Upon discontinuation of both drugs, performance progressively returned to baseline. The four alprazolam DRCs as well as the effect-time profiles demonstrated that caffeine altered neither the magnitudes nor the patterns of alprazolam effects on the two rates under chronic dose regimens. The Pöch DRC method further confirmed the independent interaction of alprazolam and caffeine. Thus, the independence of the interaction held for both the acute and chronic dose regimens despite the development of tolerance in the latter regimens.     

Management of asthma and chronic airflow limitation: Are methylxanthines obsolete?

The widespread popularity of methylxanthine derivatives should be reassessed in light of current evidence. These drugs are relatively weak bronchodilators, respiratory muscle stimulants and inotropic agents and adverse effects, sometimes life threatening, occur fairly frequently. In contrast, beta-2 adrenergic and anticholinergic bronchodilator aerosols used in asthma or chronic obstructive lung disease, and the prophylactic anti-inflammatory aerosols of corticosteroids and cromolyn provide a spectrum of therapeutic choices which address both the inflammatory and bronchoconstrictor components of acute and chronic airflow limitation. Aerosol bronchodilators, in general, are more potent, are virtually free of important side effects, and do not require costly serum level monitoring. Adrenoceptor agonists, together with inhaled steroids, should be considered first-line drugs of choice in managing patients with reversible airflow obstruction associated with asthma or COPD, while methylxanthines should be relegated to the position of third or fourth line drugs, if they are to be used at all. If they are, they should be used with great caution and close patient supervision and, even then, only if benefit, over and above the aerosol bronchodilators and inhaled anti-inflammatory agents can be demonstrated objectively.     

Effect of acute versus chronic theophylline administration on acute restraint stress-induced increase of pentylenetetrazol seizure threshold in mice

Various stressful paradigms were found to induce anticonvulsant effects in different seizure models. Methylxanthines, such as theophylline might contribute to the reduction of restraint-induced stress. Therefore, in this study the influence of acute restraint stress on pentylenetetrazol (PTZ) seizure thresholds as well as the effect of acute and chronic theophylline pretreatment on stress-induced modulation of the seizure threshold were assessed in mice. The onset of the three consecutive seizure phases: myoclonic twitch (MTW), generalized clonus (GNCL) and tonic hind limb extension (THE) was delayed after exposure to a 2 h restraint stress by 34%, 23% and 24%, respectively. In nonstressed mice, acute theophylline injection (100 mg/kg, i.p.) decreased the threshold only for THE. However, in stressed animals, the pretreatment with the methylxanthine significantly enhanced the dose of the convulsant producing the same seizure phase. In nonstressed mice, long-term theophylline treatment (50 mg/kg, twice daily for 14 days) increased PTZ threshold for all three seizure phases. In contrast, in chronically treated with theophylline mice exposed to restraint stress, significant decrease in the PTZ threshold for all seizure phases compared to control stressed animals have been observed. These results suggest that, depending on the treatment regimen (acute versus chronic), theophylline specifically and differentially modulates the anticonvulsant effect of restraint stress in mice.     

Discriminative stimulus and subjective effects of theobromine and caffeine in humans

Theobromine versus placebo discrimination and caffeine versus placebo discrimination were studied in two consecutive experiments in seven volunteers who abstained from methylxanthines. Daily sessions involved PO double-blind ingestion of two sets of capsules sequentially, one of which contained drug and the other placebo. Subjects attempted to identify, and were later informed, which set of capsules contained the drug. In each experiment subjects were exposed to progressively lower doses. Five subjects acquired the theobromine discrimination; the lowest dose discriminated ranged from 100 to 560 mg. All seven subjects acquired the caffeine discrimination; the lowest dose discriminated ranged from 1.8 to 178 mg. A final experiment evaluated subjective effect ratings following 560 mg theobromine, 178 mg caffeine and placebo, which were administered double-blind in capsules once daily, five times each in mixed sequence. Caffeine produced changes in both group and individual ratings (e.g. increased well-being, energy, social disposition and alert). Theobromine did not produce changes in group ratings but changed ratings in some subjects. Across subjects, sensitivity to caffeine discriminative effects in the discrimination experiment correlated significantly with the number and magnitude of caffeine subjective effects in the final experiment. This study documents modest discriminative effects of theobromine in humans, but the basis of the discrimination is unclear. This study suggests that commonly consumed cocoa products contain behaviorally active doses of caffeine and possibly theobromine.     

Dopamine denervation leads to an increase in the intramembrane interaction between adenosine A2 and dopamine D2 receptors in the neostriatum

We have previously found, in striatal membrane preparations from young (2 months old) rats, that stimulation of adenosine A₂ receptors (with the selective adenosine A₂ agonist CGS 21680) increases the dissociation constants of high- (K_h) and low-affinity (K_l) dopamine D₂ binding sites (labelled with the selective dopamine D₂ antagonist [³H]raclopride) without changing the proportion of high affinity binding sites (R_h). In the present study in striatal preparations from adult (6 months old) rats, it was found that in addition to the increase in both K_h and K_l values, stimulation of adenosine A₂ receptors is associated with an increase in R_h. These result suggest that, in the adult rat, adenosine A₂ stimulation may inhibit the behavioural effects induced by dopamine D₂ stimulation both by decreasing the affinity and the transduction of dopamine D₂ receptors. We have also studied the intramembrane A₂-D₂ receptor interaction in an experimental model of Parkinson's disease, namely in rats with a unilateral 6-OH-dopamine-induced lesion of the nigro-striatal dopamine pathway. It was found that a unilateral dopamine denervation is associated with a higher density of striatal dopamine D₂ receptors in the order of 20%, without any change in their affinity compared with the unlesioned neostriatum. Furthermore, the density (B_max values) of dopamine D₂ receptors in the contralateral neostriatum was significantly higher (about 20%) than in the striatum from native animals. This finding suggests that an unilateral dopamine denervation also induces compensatory long-lasting changes of dopamine D₂ receptors in the contralateral neostriatum. In addition to the hightened sensitivity to dopamine agonists, it is known that the dopamine denervated striatum is more sensitive to adenosine antagonists like methylxanthines. If the adenosine A₂-dopamine D₂ interaction is the main mechanism of action mediating the central effects of methylxanthines, the dopamine denervation might also potentiate this interaction, i.e., dopamine D₂ receptors could be not only more sensitive to dopamine but also to adenosine A₂ receptor activation. Our results support this hypothesis, since membrane preparations from the denervated neostriatum are more sensitive to the effect of CGS 21680 on dopamine D₂ receptors. Thus a low dose of CGS 21680 (3 nM), which is not effective in membrane preparations from the neostriatum of naive animals, is still effective in membranes from the denervated neostriatum. These results underline the potential antiparkinsonian activity of adenosine A₂ antagonists.     

荧光法研究3种甲基黄嘌呤类生物碱与牛血清白蛋白的相互作用

目的:研究咖啡因、茶碱、可可碱3种甲基黄嘌呤类生物碱与牛血清白蛋白(BSA)结合反应的荧光光谱行为,了解其与蛋白质结合的信息。方法:采用荧光光谱法研究25℃和37℃时3种生物碱不同浓度对BSA的荧光猝灭作用,通过Stern-Volmer曲线和Perrin曲线线性拟合计算得到动态猝灭常数和静态猝灭常数,判断猝灭机制;计算结合常数、结合位点数及热力学函数ΔH、ΔS、ΔG,判断其结合作用力模型。结果:随着3种生物碱浓度升高,BSA内源荧光强度有规律地降低;随温度的升高,静态猝灭常数和结合常数均降低,而3种生物碱与BSA的结合常数差别较大,但结合位点数均接近1;ΔG<0,ΔH<0,ΔS>0。结论:3种生物碱能较显著猝灭BSA的荧光,且机制均为静态猝灭,与白蛋白之间均存在以疏水相互作用为主的结合作用;3种结构相近的生物碱与BSA的相互作用存在差异。     

Aminophylline increases ventilation and diaphragm contractility in awake canines

Traditional theophylline bronchodilators are still used clinically, especially in COPD. However, the effect of theophyllines on ventilation and respiratory muscles remains uncertain and these effects have not been measured directly in any awake, intact mammal. We hypothesized that aminophylline in the usual therapeutic dosage range, would elicit in the awake mammal, a significant increase in ventilation, and a significant increase in costal diaphragm shortening and contractility as recorded directly from the muscle. Therefore, we studied 13 awake canines, which had been chronically implanted with fine-wire EMG electrodes and sonomicrometer crystals in the costal segment of the diaphragm. Ventilatory parameters, moving average muscle EMG activity and muscle length and shortening, were measured at baseline and with aminophylline, during resting and hypercapnic stimulated breathing. Experiments were carried out prior to administration of aminophylline (baseline), and 1.5 h after loading and ongoing infusion with aminophylline. Minute ventilation, tidal volume and respiratory frequency all increased significantly with aminophylline, both during resting breathing and at equivalent levels of hypercapnic stimulated breathing. Costal diaphragm baseline muscle length was entirely unchanged with aminophylline. Costal diaphragm shortening increased significantly with aminophylline while corresponding costal diaphragm EMG activity remained constant, consistent with increased diaphragm contractility. Thus, in awake, intact mammals, aminophylline in usual therapeutic dosage elicits increased ventilation and increased contractility of respiratory muscles.