Maggi C A, Giuliani S, Santicioli P
Pharmacology Department, A. Menarini Pharmaceuticals, Florence, Italy.
Naunyn Schmiedebergs Arch Pharmacol. 1994 May;349(5):510-22. doi: 10.1007/BF00169141.
We have investigated the effect of the dihydropyridine calcium channel agonist, Bay K 8644, and of the plant alkaloid blocker of calcium-induced calcium release (CICR) from the sarcoplasmic reticulum, ryanodine, on the refractory period, action potential and mechanical response of the guinea-pig isolated ureter to electrical stimulation. All experiments were performed in ureters pre-exposed to 10 microM capsaicin to eliminate the inhibitory influence exerted by local release of sensory neuropeptides on ureteral excitability and contraction. In organ bath experiments, electrical field stimulation with parameters which produce direct excitation of ureteral smooth muscle (train of pulses at 10 Hz, 5 ms pulse width, 60 V for 1 s) produced tetrodotoxin- (1 microM) resistant phasic contractions. The response to EFS was abolished by nifedipine (1 nM-3 microM) and was enhanced by Bay K 8644 (1 nM-3 microM). In the presence of Bay K 8644 (1 microM), nifedipine (30 microM) abolished the evoked contractions. Ryanodine (10-100 microM) had no significant effect on the amplitude of evoked contraction. The response of the guinea-pig ureter to direct electrical stimulation of smooth muscle is characterized by a refractory period: at least 40 s interstimulus interval was required to produce a second response in all preparations tested. Bay K 8644 (1 microM) markedly reduced the refractory period of the ureter and a similar effect was observed with ryanodine (100 microM). To further analyze the effect of Bay K 8644 and ryanodine on the refractory period, the response of the ureter was investigated over a 10 s period of stimulation (other parameters as above). In control ureters, continuous stimulation for 10 s produced only one phasic contraction just after the beginning of the train of stimuli. In the presence of Bay K 8644 or ryanodine, more than one phasic contraction developed during a 10 s stimulation, i.e. the refractory period became shorter than the train duration. When both Bay K 8644 and ryanodine were tested on the same preparations, an additive excitatory effect was observed on the mechanical response to electrical stimulation. A slight elevation of KCl concentration (5-10 mM) reduced the refractory period of the ureter as observed with ryanodine or Bay K 8644. Application of KCl (80 mM) produced a biphasic contractile response of the ureter: a series of phasic contractions occurred first, which were then replaced by a slowly developing tonic response. Bay K 8644 (1 microM) enhanced both components of the response to KCl. Ryanodine (10 and 100 microM) markedly prolonged the duration of phasic contractions evoked by KCl and, at 100 microM, slightly (about 25%) reduced the amplitude of tonic contraction.(ABSTRACT TRUNCATED AT 400 WORDS)
我们研究了二氢吡啶类钙通道激动剂Bay K 8644以及植物生物碱(它是肌浆网钙诱导钙释放(CICR)的阻滞剂)ryanodine对豚鼠离体输尿管电刺激后的不应期、动作电位和机械反应的影响。所有实验均在预先暴露于10微摩尔辣椒素的输尿管中进行,以消除感觉神经肽局部释放对输尿管兴奋性和收缩产生的抑制性影响。在器官浴实验中,用能直接激发输尿管平滑肌的参数进行电场刺激(10赫兹的脉冲串,5毫秒脉冲宽度,60伏,持续1秒),产生了对河豚毒素(1微摩尔)有抗性的相性收缩。硝苯地平(1纳摩尔至3微摩尔)可消除对电场刺激的反应,而Bay K 8644(1纳摩尔至3微摩尔)可增强该反应。在存在Bay K 8644(1微摩尔)的情况下,硝苯地平(30微摩尔)可消除诱发的收缩。Ryanodine(10至100微摩尔)对诱发收缩的幅度无显著影响。豚鼠输尿管对平滑肌直接电刺激的反应具有不应期特征:在所有测试标本中,至少需要40秒的刺激间隔才能产生第二次反应。Bay K 8644(1微摩尔)显著缩短了输尿管的不应期,ryanodine(100微摩尔)也观察到类似效果。为进一步分析Bay K 8644和ryanodine对不应期的影响,在10秒的刺激期内研究了输尿管的反应(其他参数如上)。在对照输尿管中,持续刺激10秒仅在刺激串开始后产生一次相性收缩。在存在Bay K 8644或ryanodine的情况下,在10秒刺激期间会产生不止一次相性收缩,即不应期变得短于刺激持续时间。当在同一标本上同时测试Bay K 8644和ryanodine时,观察到对电刺激的机械反应有相加的兴奋作用。氯化钾浓度轻微升高(5至10毫摩尔)可缩短输尿管的不应期,这与ryanodine或Bay K 8644的作用类似。应用氯化钾(80毫摩尔)可使输尿管产生双相收缩反应:首先出现一系列相性收缩,随后被缓慢发展的紧张性反应所取代。Bay K 8644(1微摩尔)增强了对氯化钾反应的两个成分。Ryanodine(10和100微摩尔)显著延长了氯化钾诱发的相性收缩的持续时间,在100微摩尔时,轻微(约25%)降低了紧张性收缩的幅度。(摘要截短于400字)