Chang L C, Wang J P
Department of Education and Research, Taichung Veterans General Hospital, Taichung, Taiwan.
Eur J Pharmacol. 2000 Feb 25;390(1-2):61-6. doi: 10.1016/s0014-2999(00)00033-9.
The signaling pathways leading to p38 mitogen-activated protein kinase (MAPK) activation in formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated rat neutrophils were examined. Immunoblot analysis with antibodies against a phosphorylated form of p38 MAPK showed that fMLP-stimulated p38 MAPK activation was dependent on a pertussis toxin-sensitive G protein. Two phosphatidylinositol 3-kinase inhibitors, wortmannin and 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002), did not affect the p38 MAPK activation. Phosphorylation of p38 MAPK was concentration dependently attenuated by a tyrosine kinase inhibitor, genistein, and by a Ca(2+)-dependent protein kinase C inhibitor, 13-cyanoethyl-12-methyl-6,7,12,13-tetrahydroindolo[2,3-a]pyrrolo[3 , 4-c]carbazole-7-one (Gö6976). However, the protein kinase C inhibitors with a broader spectrum, 2-[1-(3-dimethylaminopropyl)-5-methoxy-1H-indol-3-yl]-3-(1H-indol-3-y l)-maleimide (Gö6983) and 2-[1-(3-dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimi de (GF109203X), had no inhibitory effect. fMLP-stimulated p38 MAPK phosphorylation was also reduced in cells pretreated with a phospholipase C inhibitor, 1-[6-((17beta-3-methoxyestra-1,3, 5(10)-trien-17-yl)amino)hexyl]-1H-pyrrole-2,5-dione (U73122), or preloaded with an intracellular Ca(2+) chelator, 1, 2-bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA). We conclude that phosphorylation of p38 MAPK by fMLP stimulation in rat neutrophils is dependent on G(i/o) protein, nonreceptor tyrosine kinase, phospholipase C/Ca(2+), and probably Ca(2+)-dependent protein kinase C pathways.
研究了在甲酰甲硫氨酰亮氨酰苯丙氨酸（fMLP）刺激的大鼠中性粒细胞中导致p38丝裂原活化蛋白激酶（MAPK）激活的信号通路。用针对p38 MAPK磷酸化形式的抗体进行免疫印迹分析表明，fMLP刺激的p38 MAPK激活依赖于百日咳毒素敏感的G蛋白。两种磷脂酰肌醇3激酶抑制剂渥曼青霉素和2-（4-吗啉基）-8-苯基-4H-1-苯并吡喃-4-酮（LY294002）不影响p38 MAPK的激活。酪氨酸激酶抑制剂染料木黄酮和钙依赖性蛋白激酶C抑制剂13-氰基乙基-12-甲基-6,7,12,13-四氢吲哚并[2,3-a]吡咯并[3,4-c]咔唑-7-酮（Gö6976）可浓度依赖性地减弱p38 MAPK的磷酸化。然而，具有更广泛作用谱的蛋白激酶C抑制剂2-[1-（3-二甲基氨基丙基）-5-甲氧基-1H-吲哚-3-基]-3-（1H-吲哚-3-基）-马来酰亚胺（Gö6983）和2-[1-（3-二甲基氨基丙基）-1H-吲哚-3-基]-3-（1H-吲哚-3-基）-马来酰亚胺（GF109203X）没有抑制作用。在用磷脂酶C抑制剂1-[6-（（17β-3-甲氧基雌甾-1,3,5（10）-三烯-17-基）氨基）己基]-1H-吡咯-2,5-二酮（U73122）预处理的细胞中，或预先加载细胞内钙螯合剂1,2-双-（邻氨基苯氧基）乙烷-N,N,N',N'-四乙酸（BAPTA）的细胞中，fMLP刺激的p38 MAPK磷酸化也降低。我们得出结论，fMLP刺激大鼠中性粒细胞中p38 MAPK的磷酸化依赖于G（i/o）蛋白、非受体酪氨酸激酶、磷脂酶C/钙和可能的钙依赖性蛋白激酶C途径。
