Jarajapu Yagna P R, Knot Harm J
Department of Pharmacology and Therapeutics and McKnight Brain Institute, Vascular Biology and Cell Physiology Group, University of Florida College of Medicine, Gainesville 32610-0267, USA.
Am J Physiol Heart Circ Physiol. 2002 Dec;283(6):H2234-8. doi: 10.1152/ajpheart.00624.2002. Epub 2002 Aug 29.
Earlier studies have implicated phospholipase C (PLC) in the development of myogenic tone (MT) based on pharmacological studies in larger arteries. In the present study, we further investigated the cellular effects of PLC inhibition using pharmacological and electrophysiological approaches to provide more quantitative functional evidence for the involvement of PLC in the genesis of MT in small cerebral arteries. The phosphatidylinositol-selective PLC (PI-PLC) inhibitor U-73122 decreased MT by 87% in posterior cerebral arteries from Sprague-Dawley rats with pIC(50) of 6.2 +/- 0.09 (n = 5). Similar potency (pIC(50) of 6.2 +/- 0.04, n = 5) was observed in arteries with MT that were further constricted with 30 nM serotonin. The phosphatidylcholine-specific (PC-PLC) inhibitor D609 had no effect on MT. U-73343, the inactive analog of U-73122, did not show any relaxant effect, but at higher concentrations (>1 microM) it reduced MT. In the presence of 125-500 nM U-73122, the pressure-diameter curves shifted toward that obtained in Ca-free conditions. U-73122-mediated decrease in MT was accompanied by a decrease in mean arterial wall calcium (maximum effect: 77 +/- 3% of 16 mM KCl-mediated decrease, n = 4). This was due to a simultaneous membrane potential hyperpolarization of approximately 9 mV or from -44 +/- 1 to -53 +/- 2 mV (10 microM, P < 0.001, n = 8). In summary, this study provides the first quantitative data suggesting a critical importance of PI-PLC in the genesis of pressure-induced MT in rat cerebral arteries via membrane potential depolarization and increased calcium influx.
早期研究基于对较大动脉的药理学研究,表明磷脂酶C(PLC)参与肌源性张力(MT)的形成。在本研究中,我们使用药理学和电生理学方法进一步研究了PLC抑制的细胞效应,以提供更定量的功能证据,证明PLC参与小鼠脑动脉MT的发生。磷脂酰肌醇选择性PLC(PI-PLC)抑制剂U-73122使来自Sprague-Dawley大鼠的大脑后动脉中的MT降低了87%,其pIC(50)为6.2±0.09(n = 5)。在用30 nM血清素进一步收缩的具有MT的动脉中观察到了类似的效力(pIC(50)为6.2±0.04,n = 5)。磷脂酰胆碱特异性(PC-PLC)抑制剂D609对MT没有影响。U-73122的无活性类似物U-73343没有显示出任何舒张作用,但在较高浓度(>1 microM)时它降低了MT。在存在125 - 500 nM U-73122的情况下,压力-直径曲线向在无钙条件下获得的曲线移动。U-73122介导的MT降低伴随着平均动脉壁钙的减少(最大效应:为16 mM KCl介导的减少的77±3%,n = 4)。这是由于同时发生了约9 mV的膜电位超极化,从-44±1 mV变为-53±2 mV(10 microM,P < 0.001,n = 8)。总之,本研究提供了首个定量数据,表明PI-PLC在大鼠脑动脉压力诱导的MT形成中通过膜电位去极化和钙内流增加起着至关重要的作用。