Abstract:
Basophils and mast cells are specialized effector cells in allergic reactions. Haliotis discus hannai (abalone), is valuable seafood. Abalone male viscera, which has a brownish color and has not been previously reported to show anti-allergic activities, was extracted with acetone. Six different acetone/hexane fractions (0, 10, 20, 30, 40, and 100%) were obtained using a silica column via β-hexosaminidase release inhibitory activity-guided selection in phorbol myristate acetate and a calcium ionophore, A23187 (PMACI)-induced human basophils, KU812F cells. The 40% acetone/hexane fraction (A40) exhibited the strongest inhibition of PMACI-induced-β-hexosaminidase release. This fraction dose-dependently inhibited reactive oxygen species (ROS) production and calcium mobilization without cytotoxicity. Western blot analysis revealed that A40 down-regulated PMACI-induced MAPK (ERK 1/2, p-38, and JNK) phosphorylation, and the NF-κB translocation from the cytosol to membrane. Moreover, A40 inhibited PMACI-induced interleukin (IL)-1β, IL-6, and IL-8 production. Anti-allergic activities of A40 were confirmed based on inhibitory effects on IL-4 and tumor necrosis factor alpha (TNF-α) production in compound (com) 48/80-induced rat basophilic leukemia (RBL)-2H3 cells. A40 inhibited β-hexosaminidase release and cytokine production such as IL-4 and TNF-α produced by com 48/80-stimulated RBL-2H3 cells. Furthermore, it\'s fraction attenuated the IgE/DNP-induced passive cutaneous anaphylaxis (PCA) reaction in the ears of BALB/c mice. Our results suggest that abalone contains the active fraction, A40 is a potent therapeutic and functional material to treat allergic diseases.
摘要:
嗜碱性粒细胞和肥大细胞是过敏反应中的特化效应细胞。Haliotisdiscushannai(鲍鱼),是珍贵的海鲜。鲍鱼雄性内脏,呈褐色,以前没有报道显示抗过敏活性,用丙酮提取。使用二氧化硅柱,通过β-己糖胺酶释放抑制活性指导选择佛波醇肉豆蔻酸酯乙酸酯和钙离子载体,获得了六种不同的丙酮/己烷级分(0、10、20、30、40和100%),A23187(PMACI)诱导的人类嗜碱性粒细胞,KU812F细胞。40%丙酮/己烷级分(A40)对PMACI诱导的β-己糖胺酶释放的抑制作用最强。该部分剂量依赖性地抑制了活性氧(ROS)的产生和钙的动员,而没有细胞毒性。蛋白质印迹分析显示A40下调PMACI诱导的MAPK(ERK1/2、p-38和JNK)磷酸化,和NF-κB从细胞质易位到膜。此外,A40抑制PMACI诱导的白细胞介素(IL)-1β,IL-6和IL-8的产生。基于对化合物(com)48/80诱导的大鼠嗜碱性粒细胞白血病(RBL)-2H3细胞中IL-4和肿瘤坏死因子α(TNF-α)产生的抑制作用,证实了A40的抗过敏活性。A40抑制β-己糖胺酶的释放和细胞因子的产生,例如由com48/80刺激的RBL-2H3细胞产生的IL-4和TNF-α。此外,它的分数减弱了BALB/c小鼠耳朵中IgE/DNP诱导的被动皮肤过敏反应(PCA)。我们的结果表明,鲍鱼含有活性部分,A40是一种有效的治疗和功能性材料,用于治疗过敏性疾病。