Kawaguchi A, Ohmori M, Harada K, Tsuruoka S, Sugimoto K, Fujimura A
Department of Clinical Pharmacology, Jichi Medical School, 3311-1 Minamikawachi-machi, Kawachi-gun, 329-0498, Tochigi, Japan.
Eur J Pharmacol. 2000 Sep 8;403(3):203-8. doi: 10.1016/s0014-2999(00)00593-8.
We investigated the involvement of p160ROCK (a Rho-associated coiled coil-forming protein kinase), one of Rho kinases on superoxide anion production (O(2)(-) production), aggregation and adhesion of human polymorphonuclear leukocytes under physiological condition, using a selective p160ROCK inhibitor, (+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide (Y-27632). Y-27632 inhibited the O(2)(-) production stimulated by phorbol-12-myristate-13-acetate (PMA) in a dose-dependent manner. Stauroprorine blocked the PMA-induced O(2)(-) production while wortmannin did not. Y-27632 also inhibited the O(2)(-) production by guanosine 5'-O-(3-thiotriphosphate) (GTP(gamma)S) 100 microM. N-formyl-Met-Leu-Phe (fMLP)-induced O(2)(-) production was not influenced by Y-27632, but was inhibited by wortmannin. The enhanced O(2)(-) production by Ca-ionophore A23817 and thapsigargin was not inhibited by Y-27632. Y-27632 did not change the basal intracellular Ca(2+) concentration nor its elevation stimulated by fMLP. Polymorphonuclear leukocytes aggregation induced by PMA was dose-dependently decreased by Y-27632 while their aggregation stimulated by fMLP was enhanced by the agent. Polymorphonuclear leukocytes adhesion induced by PMA or fMLP was not influenced by Y-27632.These results suggest that p160ROCK is involved in the PMA-induced O(2)(-) production and aggregation in human polymorphonuclear leukocytes. This kinase might locate in downstream of protein kinase C in polymorphonuclear leukocytes.
我们使用一种选择性p160ROCK抑制剂（+）-（R）-反式-4-（1-氨乙基）-N-（4-吡啶基）环己烷甲酰胺（Y-27632），研究了Rho激酶之一p160ROCK在生理条件下对人多形核白细胞超氧阴离子产生（O₂⁻产生）、聚集和黏附的影响。Y-27632以剂量依赖性方式抑制佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）刺激的O₂⁻产生。星形孢菌素可阻断PMA诱导的O₂⁻产生，而渥曼青霉素则不能。Y-27632还可抑制100微摩尔鸟苷5'-O-（3-硫代三磷酸）（GTPγS）诱导的O₂⁻产生。N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸（fMLP）诱导的O₂⁻产生不受Y-27632影响，但可被渥曼青霉素抑制。钙离子载体A23817和毒胡萝卜素增强的O₂⁻产生不受Y-27632抑制。Y-27632既不改变基础细胞内Ca²⁺浓度，也不改变fMLP刺激引起的Ca²⁺浓度升高。Y-27632可剂量依赖性降低PMA诱导的多形核白细胞聚集，而fMLP刺激的多形核白细胞聚集则被该药物增强。PMA或fMLP诱导的多形核白细胞黏附不受Y-27632影响。这些结果表明，p160ROCK参与了人多形核白细胞中PMA诱导的O₂⁻产生和聚集。该激酶可能位于多形核白细胞中蛋白激酶C的下游。
