Martinez-Losa M, Cortijo J, Piqueras L, Sanz M J, Morcillo E J
Departamento de Farmacologia, Facultad de Medicina, Universidad de Valencia, Valencia, Spain.
Clin Exp Allergy. 2009 Apr;39(4):537-46. doi: 10.1111/j.1365-2222.2009.03204.x. Epub 2009 Feb 13.
Eosinophils are prominent effectors of allergic inflammation. Taurine-chloramine (TauCl), a derivative of the amino acid taurine, shows antioxidant properties in different cell systems but its effects on eosinophils have not been reported.
To study the effects of TauCl and taurine on functional responses of isolated human eosinophils activated by different stimuli.
Human eosinophils were purified from the blood of healthy donors by a magnetic bead separation system. The effects of TauCl and taurine (0.1-1 mM) were investigated on the generation of superoxide anion (ferricytochrome-c reduction microassay), calcium signal (fluorimetry), p47phox-p67phox translocation (Western blot), leukotriene C4 (LTC4) production (enzymeimmunoassay), eosinophil peroxidase (EPO) release (spectrophotometry), eosinophil cationic protein (ECP) release (radioimmunoassay), apoptosis (flow cytometry with annexin V-propidium iodide), and nuclear factor-kappaB (NF-kappaB) activation (Western blot).
TauCl inhibited superoxide anion generation triggered by N-formyl-Met-Leu-Phe (fMLP; 30 nM), phorbol myristate acetate (1 nM) and serum opsonized zymosan (0.5 mg/mL) with similar potency (IC50 approximately 200 microM) for the three stimuli, while taurine (0.1-1 mM) was scarcely effective. TauCl but not taurine inhibited p47phox-p67phox translocation. TauCl (200 microM) and taurine (1 mM) did not modify the [Ca2+]i responses to fMLP. TauCl inhibited the release of EPO (IC50 approximately 200 microM) and reduced ECP and LTC4 production from fMLP-activated eosinophils while taurine was without significant effects. TauCl (1 mM) did not change constitutive apoptosis but significantly attenuated the ability of granulocyte-monocyte colony-stimulating factor (GM-CSF) and IL-5 to prevent apoptosis. The activation of eosinophil NF-kappaB induced by GM-CSF and IL-5 was suppressed by TauCl.
Taurine is without significant in vitro effects on human eosinophil functions but its derivative TauCl inhibits oxidative burst and generation of inflammatory mediators, and reverses the survival effect produced by inflammatory cytokines. Therefore, endogenous TauCl may help to suppress excessive inflammatory response in eosinophils at inflammatory sites.
嗜酸性粒细胞是过敏性炎症的主要效应细胞。牛磺酸氯胺(TauCl)是氨基酸牛磺酸的衍生物,在不同细胞系统中表现出抗氧化特性,但其对嗜酸性粒细胞的影响尚未见报道。
研究TauCl和牛磺酸对不同刺激激活的人离体嗜酸性粒细胞功能反应的影响。
通过磁珠分离系统从健康供者血液中纯化人嗜酸性粒细胞。研究了TauCl和牛磺酸(0.1 - 1 mM)对超氧阴离子生成(高铁细胞色素c还原微量测定法)、钙信号(荧光测定法)、p47phox - p67phox转位(蛋白质免疫印迹法)、白三烯C4(LTC4)生成(酶免疫测定法)、嗜酸性粒细胞过氧化物酶(EPO)释放(分光光度法)、嗜酸性粒细胞阳离子蛋白(ECP)释放(放射免疫测定法)、凋亡(膜联蛋白V - 碘化丙啶流式细胞术)以及核因子 - κB(NF - κB)激活(蛋白质免疫印迹法)的影响。
TauCl抑制由N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸(fMLP;30 nM)、佛波酯(1 nM)和血清调理酵母聚糖(0.5 mg/mL)触发的超氧阴离子生成,对这三种刺激的抑制效力相似(IC50约为200 μM),而牛磺酸(0.1 - 1 mM)几乎无效。TauCl而非牛磺酸抑制p47phox - p67phox转位。TauCl(200 μM)和牛磺酸(1 mM)不改变对fMLP的[Ca2 + ]i反应。TauCl抑制EPO释放(IC50约为200 μM),并减少fMLP激活嗜酸性粒细胞产生的ECP和LTC4,而牛磺酸无显著作用。TauCl(1 mM)不改变基础凋亡,但显著减弱粒细胞 - 单核细胞集落刺激因子(GM - CSF)和IL - 5预防凋亡的能力。TauCl抑制GM - CSF和IL - 5诱导的嗜酸性粒细胞NF - κB激活。
牛磺酸在体外对人嗜酸性粒细胞功能无显著影响,但其衍生物TauCl抑制氧化爆发和炎症介质生成,并逆转炎症细胞因子产生的存活效应。因此,内源性TauCl可能有助于抑制炎症部位嗜酸性粒细胞的过度炎症反应。