Kawakami Ayako, Suzukawa Maho, Koketsu Rikiya, Komiya Akiko, Ohta Ken, Yamamoto Kazuhiko, Yamaguchi Masao
Department of Allergy and Rheumatology, University of Tokyo Graduate School of Medicine, Tokyo, Japan.
Allergy Asthma Proc. 2008 May-Jun;29(3):322-8. doi: 10.2500/aap.2008.29.3118.
Regulation of basophil survival is an important aspect in the pathogenesis of allergic inflammation associated with local accumulation of basophils. However, pharmacologic modulation of basophil survival is largely unknown except for the apoptosis-enhancing effect of glucocorticoids. We tested the effects of two anti-allergic and anti-asthmatic drugs, olopatadine and theophylline, on basophil survival. Basophils were highly purified from normal human peripheral blood. Apoptosis was analyzed by flow cytometry using annexin V staining or another staining method that detected alterations in the mitochondrial transmembrane potential. In addition to the conventional method using annexin V, basophil apoptosis was successfully established by analysis of the mitochondrial transmembrane potential. Olopatadine decreased the number of live basophils, and they induced apoptosis of basophils during culture. The decline in live basophils was induced by olopatadine even when low doses of IL-3 were included in the culture medium. Theophylline also affected basophil apoptosis and induced a decrease in the number of live basophils. Basophil apoptosis was enhanced by both olopatadine and theophylline. This effect may partly explain the pharmacologic basis of why these drugs are effective on allergic diseases.
嗜碱性粒细胞存活的调节是与嗜碱性粒细胞局部聚集相关的过敏性炎症发病机制中的一个重要方面。然而,除了糖皮质激素的促凋亡作用外,关于嗜碱性粒细胞存活的药物调节作用在很大程度上尚不清楚。我们测试了两种抗过敏和抗哮喘药物奥洛他定和茶碱对嗜碱性粒细胞存活的影响。嗜碱性粒细胞从正常人外周血中高度纯化得到。使用膜联蛋白V染色或另一种检测线粒体跨膜电位变化的染色方法,通过流式细胞术分析细胞凋亡。除了使用膜联蛋白V的传统方法外,通过分析线粒体跨膜电位成功建立了嗜碱性粒细胞凋亡模型。奥洛他定减少了活嗜碱性粒细胞的数量,并且在培养过程中诱导嗜碱性粒细胞凋亡。即使在培养基中加入低剂量的白细胞介素-3,奥洛他定也能诱导活嗜碱性粒细胞数量减少。茶碱也影响嗜碱性粒细胞凋亡,并导致活嗜碱性粒细胞数量减少。奥洛他定和茶碱均增强了嗜碱性粒细胞凋亡。这种作用可能部分解释了这些药物对过敏性疾病有效的药理学基础。