Gupte Sachin A, Arshad Muhammad, Viola Steven, Kaminski Pawel M, Ungvari Zoltan, Rabbani Golam, Koller Akos, Wolin Michael S
Department of Physiology, New York Medical College, Valhalla, NY 10595, USA.
Am J Physiol Heart Circ Physiol. 2003 Dec;285(6):H2316-26. doi: 10.1152/ajpheart.00229.2003. Epub 2003 Aug 21.
Pentose phosphate pathway (PPP) inhibitors, 6-aminonicotinamide (6-AN) and epiandrosterone (Epi), were employed to examine whether changes in NADP(H) redox regulates contractile force in endothelium-removed bovine coronary arteries (BCAs). 6-AN (0.01-5 mM) or Epi (1-500 microM) elicited dose-dependent relaxation in BCAs contracted with 30 mM KCl, 0.1 microM U-44619, and endothelin-1 but not with phorbol 12,13-dibutyrate, a protein kinase C activator that causes Ca2+-independent contraction. Relaxation to PPP inhibition was associated with oxidation of NADPH and glutathione (GSH). Relaxation to 6-AN was not mediated by H2O2, because it was not altered by hypoxia or the peroxide scavenger ebselen (100 microM). The thiol reductant DTT (3 mM) attenuated the relaxation to 6-AN and Epi by 30-40%. Inhibition of glycolysis or mitochondrial electron transport did not elicit relaxation in BCAs contracted with 30 mM KCl, suggesting these pathways may not be involved in relaxation elicited by PPP inhibition. High doses of K+ channel blockers [e.g., TEA (10 mM) and 4-aminopyridine (10 mM)] only partially inhibited the relaxation to 6-AN. On the basis of changes in the fura-2 fluorescence ratio, 6-AN and Epi appeared to markedly reduce intracellular Ca2+. Thus PPP inhibition oxidizes NADPH and GSH and appears to activate a novel coordination of redox-controlled relaxing mechanisms in BCAs mediated primarily through decreasing intracellular Ca2+.
采用戊糖磷酸途径(PPP)抑制剂6-氨基烟酰胺(6-AN)和表雄酮(Epi),研究NADP(H)氧化还原变化是否调节去内皮牛冠状动脉(BCA)的收缩力。6-AN(0.01 - 5 mM)或Epi(1 - 500 μM)可使由30 mM KCl、0.1 μM U-44619和内皮素-1引起收缩的BCA产生剂量依赖性舒张,但对佛波醇12,13 - 二丁酸(一种引起非钙依赖性收缩的蛋白激酶C激活剂)引起收缩的BCA无此作用。对PPP抑制的舒张作用与NADPH和谷胱甘肽(GSH)的氧化有关。对6-AN的舒张作用不是由H2O2介导的,因为缺氧或过氧化物清除剂依布硒啉(100 μM)对其无影响。硫醇还原剂二硫苏糖醇(DTT,3 mM)使对6-AN和Epi的舒张作用减弱30 - 40%。抑制糖酵解或线粒体电子传递未引起由30 mM KCl引起收缩的BCA舒张,提示这些途径可能不参与PPP抑制引起的舒张。高剂量的钾通道阻滞剂[如四乙铵(TEA,10 mM)和4-氨基吡啶(10 mM)]仅部分抑制对6-AN的舒张作用。根据fura-2荧光比率的变化,6-AN和Epi似乎能显著降低细胞内Ca2+。因此,PPP抑制可氧化NADPH和GSH,并似乎激活了一种新的氧化还原控制的舒张机制协同作用,在BCA中主要通过降低细胞内Ca2+介导。