Maggi C A, Giuliani S, Santicioli P
Pharmacology Department A. Menarini Pharmaceuticals, Florence, Italy.
Naunyn Schmiedebergs Arch Pharmacol. 1995 Nov;352(5):529-39. doi: 10.1007/BF00169387.
We aimed at studying the mechanism(s) of the inhibitory effect exerted by calcitonin gene-related peptide (CGRP) on the spontaneous activity of the guinea-pig isolated renal pelvis. In organ bath experiments, CGRP (1-100 nM) produced a concentration-dependent (EC50 8 nM) partial inhibition (Emax about 35% inhibition of motility index) of spontaneous contractions. The potassium (K) channel opener, cromakalim (3-10 microM) promptly suppressed the spontaneous contractions in a glibenclamide-(10 microM) sensitive manner. Glibenclamide (10 microM) did not affect the inhibitory action of CGRP. The calcium (Ca) channel agonist, Bay K 8644 (1 microM), markedly enhanced the spontaneous activity of the renal pelvis and reduced the inhibitory effect of CGRP. The protein kinase A inhibitors Rp-cAMPS (300 microM), H8 (100 microM) and H89 (10 microM), and the blockers of intracellular Ca handling by sarcoplasmic reticulum, ryanodine (100 microM) and thapsigargin (1 microM) did not affect the response to CGRP. The response to CGRP was likewise unaffected by the nitric oxide synthase inhibitor, L-nitroarginine (30 microM) and by the protein kinase G inhibitor, KT5823 (3 microM). Furthermore, the inhibitory action of CGRP was not modified by lowering the extracellular concentration of K (from 5.9 to 1.2 mM) nor by increasing (from 2.5 to 3.75 mM) or decreasing (from 2.5 to 0.25 mM) the extracellular Ca concentration. Replacement of 80% glucose with 2-deoxyglucose (2-DOG) reduced the amplitude of spontaneous contractions, both in the absence and presence of 10 microM glibenclamide. In the presence of 2-DOG, the inhibitory action of CGRP was enhanced at a similar extent, either in the absence or presence of glibenclamide. In sucrose gap, the effect of CGRP (0.1 microM for 5 min) was separately analyzed in the proximal (close to the kidney) and distal (close to the ureter) regions of the renal pelvis. Both preparations discharged spontaneous (pacemaker) action potentials having different shape, duration and frequently. CGRP had no effect on pacemaker potentials in the proximal renal pelvis while producing about 30% reduction of the frequency of pacemaker potentials and motility index in the distal renal pelvis. Cromakalim (3 microM) abolished pacemaker potentials in both regions of the renal pelvis. In conjunction with the results of previous studies in the guinea-pig ureter, the present findings document the existence of remarkable regional differences in the effector mechanisms initiated by CGRP receptor occupancy in the guinea-pig pyeloureteral tract. CGRP appears to be inherently unable to activate glibenclamide-sensitive K channels in the guinea-pig renal pelvis, a mechanism which is central for its ability to suppress latent pacemakers in the ureter. Within the renal pelvis, the sensitivity to the inhibitory effect of CGRP appears in the more distal region, from which an 'ureter-like' action potential is recorded.
我们旨在研究降钙素基因相关肽(CGRP)对豚鼠离体肾盂自发活动产生抑制作用的机制。在器官浴实验中,CGRP(1 - 100 nM)对自发收缩产生浓度依赖性(半数有效浓度8 nM)的部分抑制作用(最大效应约为运动指数抑制35%)。钾(K)通道开放剂克罗卡林(3 - 10 μM)能迅速以格列本脲(10 μM)敏感的方式抑制自发收缩。格列本脲(10 μM)不影响CGRP的抑制作用。钙(Ca)通道激动剂Bay K 8644(1 μM)显著增强肾盂的自发活动并降低CGRP的抑制作用。蛋白激酶A抑制剂Rp - cAMPS(300 μM)、H8(100 μM)和H89(10 μM),以及肌浆网对细胞内Ca处理的阻滞剂ryanodine(100 μM)和毒胡萝卜素(1 μM)均不影响对CGRP的反应。一氧化氮合酶抑制剂L - 硝基精氨酸(30 μM)和蛋白激酶G抑制剂KT5823(3 μM)同样不影响对CGRP的反应。此外,降低细胞外K浓度(从5.9 mM降至1.2 mM),或升高(从2.5 mM升至3.75 mM)或降低(从2.5 mM降至0.25 mM)细胞外Ca浓度,均不改变CGRP的抑制作用。用2 - 脱氧葡萄糖(2 - DOG)替代80%的葡萄糖可降低自发收缩的幅度,无论是否存在10 μM格列本脲。在存在2 - DOG的情况下,无论是否存在格列本脲,CGRP的抑制作用均以相似程度增强。在蔗糖间隙实验中,分别在肾盂的近端(靠近肾脏)和远端(靠近输尿管)区域分析了CGRP(0.1 μM,作用5分钟)的作用。两种标本均发放形状、持续时间和频率不同的自发(起搏)动作电位。CGRP对近端肾盂的起搏电位无影响,而使远端肾盂的起搏电位频率和运动指数降低约30%。克罗卡林(3 μM)消除了肾盂两个区域的起搏电位。结合先前在豚鼠输尿管的研究结果,本研究结果证明了豚鼠肾盂输尿管系统中CGRP受体被占据后启动的效应机制存在显著的区域差异。CGRP似乎天生无法激活豚鼠肾盂中格列本脲敏感的K通道,而该机制是其抑制输尿管潜在起搏点能力的核心。在肾盂内,对CGRP抑制作用的敏感性出现在更远端区域,从该区域可记录到“输尿管样”动作电位。