Rider L G, Hirasawa N, Santini F, Beaven M A
Arthritis and Rheumatism Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, Bethesda, MD 20892, USA.
J Immunol. 1996 Sep 15;157(6):2374-80.
Antigen stimulation of mast cells via the IgE receptor, Fc epsilon RI, results in recruitment of the cytosolic tyrosine kinases, Lyn and Syk, and the phosphorylation of proteins. We examined the effects of the glucocorticoid dexamethasone on these events in a cultured (RBL-2H3) mast cell line. Nanomolar concentrations of dexamethasone suppressed phosphorylation of proteins that were associated with the activation of the mitogen-activated protein (MAP) kinase/phospholipase A2 pathway without inhibiting initial events. For example, tyrosine phosphorylation of the subunits of Fc epsilon RI, Lyn, or Syk or of the Ras-guanine nucleotide exchange factor, Vav, was not suppressed in cells treated with up to 1 microM dexamethasone. In contrast, phosphorylation of Raf1, MEK1, p42mapk, and cytosolic phospholipase A2, as well as the associated increase in MAP kinase activity and release of arachidonic acid, were markedly inhibited in cells treated with as little as 10 nM dexamethasone--a concentration that only partially inhibited hydrolysis of inositol phospholipids or release of secretory granules. Prolonged exposure to dexamethasone also resulted in a partial decrease in expression of MEK1, p42mapk, and cytosolic phospholipase A2, which may contribute further to the effects of dexamethasone on this pathway. Activation of the MAP kinase/phospholipase A2 pathway by the calcium-mobilizing agent thapsigargin was similarly suppressed in dexamethasone-treated cells. These findings suggested that an early step in the pathway, possibly a step immediately before the activation of Raf1, was suppressed by low concentrations of dexamethasone.
经由IgE受体FcεRI对肥大细胞进行抗原刺激,会导致胞质酪氨酸激酶Lyn和Syk的募集以及蛋白质的磷酸化。我们在培养的(RBL-2H3)肥大细胞系中研究了糖皮质激素地塞米松对这些事件的影响。纳摩尔浓度的地塞米松可抑制与丝裂原活化蛋白(MAP)激酶/磷脂酶A2途径激活相关的蛋白质磷酸化,而不抑制初始事件。例如,在使用高达1μM地塞米松处理的细胞中,FcεRI、Lyn或Syk亚基或Ras-鸟嘌呤核苷酸交换因子Vav的酪氨酸磷酸化并未受到抑制。相比之下,在使用低至10 nM地塞米松处理的细胞中,Raf1、MEK1、p42mapk和胞质磷脂酶A2的磷酸化以及MAP激酶活性的相关增加和花生四烯酸的释放均受到显著抑制,该浓度仅部分抑制肌醇磷脂的水解或分泌颗粒的释放。长时间暴露于地塞米松还导致MEK1、p42mapk和胞质磷脂酶A2的表达部分下降,这可能进一步促成地塞米松对该途径的影响。在经地塞米松处理的细胞中,钙动员剂毒胡萝卜素对MAP激酶/磷脂酶A2途径的激活同样受到抑制。这些发现表明,该途径中的一个早期步骤,可能是Raf1激活之前的一个步骤,受到低浓度地塞米松的抑制。