In smooth muscle isolated from the guinea-pig stomach, cyanide (CN) and iodoacetic acid (IAA) were applied to block oxidative phosphorylation and glycolysis, respectively. Effects of IAA on generation of spontaneous mechanical and electrical activities were systematically investigated by comparing those of CN. Spontaneous activity ceased in 10-20 min during applications of 1 mM IAA. On the other hand, application of 1 mM CN also reduced the spontaneous activity, but never terminated it. In the presence of CN the negativity of the resting membrane potential was slightly reduced. 2. When spontaneous activity ceased with IAA, the resting membrane potential was not significantly affected. Also, before ceasing, the amplitude and duration of the spontaneous electrical activity were significantly reduced. The amplitude of the electrotonic potential was, however, not changed by IAA. Further, glibenclamide did not prevent the effects of IAA. These results suggest that, unlike cardiac muscle, activation of metabolism-dependent K+ channels in stomach smooth muscle does not seem to play a major role in reducing and terminating spontaneous activity during metabolic inhibition. 3. Carbachol-induced contraction transiently increased, and subsequently decreased gradually during application of IAA. 4. After 50 min application of IAA, when there was no spontaneous activity, the concentrations of phosphocreatine (PCr) and ATP measured with 31P nuclear magnetic resonance decreased to 60 and 80% of the control, respectively, while inorganic phosphate (Pi) concentration paradoxically fell to below detectable levels. During subsequent prolonged application of IAA, high-energy phosphates steadily decreased. On the other hand, after 50 min CN application, [PCr] and [ATP] decreased to approximately 30 and 80% of the control, respectively, while [Pi] increased by 2.6-fold. 5. In the presence of either CN or IAA, spontaneous mechanical and electrical activities were reduced or eliminated, although amounts of high-energy phosphates sufficient to contract smooth muscle remained. It can be postulated that some mechanism(s) related to energy metabolism, but not including ATP-sensitive K+ channels, plays an important role in generating spontaneous activity in guinea-pig stomach smooth muscle. During metabolic inhibition the energy metabolism-dependent mechanism(s) would preserve high-energy phosphates, and consequently cell viability, by stopping spontaneous activity.
摘要
在从豚鼠胃分离出的平滑肌中,分别应用氰化物(CN)和碘乙酸(IAA)来阻断氧化磷酸化和糖酵解。通过比较CN的作用,系统研究了IAA对自发机械和电活动产生的影响。在应用1 mM IAA期间,自发活动在10 - 20分钟内停止。另一方面,应用1 mM CN也降低了自发活动,但从未使其终止。在存在CN的情况下,静息膜电位的负值略有降低。2. 当IAA使自发活动停止时,静息膜电位未受到显著影响。而且,在停止之前,自发电活动的幅度和持续时间显著降低。然而,电紧张电位的幅度未被IAA改变。此外,格列本脲不能阻止IAA的作用。这些结果表明,与心肌不同,胃平滑肌中代谢依赖性钾通道的激活在代谢抑制期间减少和终止自发活动方面似乎不发挥主要作用。3. 在应用IAA期间,卡巴胆碱诱导的收缩短暂增加,随后逐渐降低。4. 在应用IAA 50分钟后,当没有自发活动时,用31P核磁共振测量的磷酸肌酸(PCr)和ATP浓度分别降至对照的60%和80%,而无机磷酸盐(Pi)浓度却反常地降至可检测水平以下。在随后长时间应用IAA期间,高能磷酸盐持续下降。另一方面,在应用CN 50分钟后,[PCr]和[ATP]分别降至对照的约30%和80%,而[Pi]增加了2.6倍。5. 在存在CN或IAA的情况下,自发机械和电活动减少或消除,尽管仍有足以使平滑肌收缩的高能磷酸盐量。可以推测,一些与能量代谢相关但不包括ATP敏感性钾通道的机制在豚鼠胃平滑肌产生自发活动中起重要作用。在代谢抑制期间,能量代谢依赖性机制将通过停止自发活动来保存高能磷酸盐,从而维持细胞活力。