山奈酚对 C57BL/6 小鼠和 LAD2 细胞 IgE 介导的过敏反应的影响。
Effect of kaempferol on IgE-mediated anaphylaxis in C57BL/6 mice and LAD2 cells.
机构信息
School of Pharmacy, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, China; Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
School of Pharmacy, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, China.
出版信息
Phytomedicine. 2020 Dec;79:153346. doi: 10.1016/j.phymed.2020.153346. Epub 2020 Sep 19.
BACKGROUND
Immunoglobulin E (IgE)-mediated mast cell (MC) activation is crucial in multiple allergic diseases. Parkinson disease protein 7 (DJ-1) and Lyn kinase were reported as the receptor-proximal events in IgE receptor (FcεRI) signals in human MC. Kaempferol, a natural flavonol mainly derived from the rhizome of traditional Chinese herb Kaempferia galanga L. (Zingiberaceae), has been known to inhibit allergic reactions, but it was limited to the receptor-distal signals on rat basophilic leukemia cells. A thorough investigation of the inhibitory effects of kaempferol on human MC has not been done.
PURPOSE
To investigate the inhibitory effects of kaempferol on IgE-mediated anaphylaxis in vivo and in human MCs, as well as the mechanism underlying its effects, especially the receptor-proximal signals.
METHODS
IgE-mediated passive cutaneous anaphylaxis and systemic anaphylaxis model were applied to elucidate the antiallergic activity of kaempferol in vivo. The degranulation assay, calcium imaging, the release of cytokines and chemokines on the laboratory of allergic disease 2 (LAD2) cells were used to evaluate the antiallergic effect of kaempferol in vitro. Western blot analysis was performed to investigate the DJ-1/Lyn signaling pathway and downstream molecules. Kinase activity assay, immunofluorescence, and molecular docking were conducted to confirm the influence of kaempferol on DJ-1/Lyn molecules.
RESULTS
Kaempferol dose-dependently attenuated ovalbumin/IgE-induced mice paw swelling, primary MC activation from paw skin, as well as rehabilitated the hypothermia, and reduced the serum concentrations of histamine, tumor necrosis factor-alpha, interleukin-8, and monocyte chemo-attractant protein-1. Additionally, kaempferol suppressed IgE-mediated LAD2 cell degranulation and calcium fluctuation. Remarkably, kaempferol was found to bind with DJ-1 protein, and initially prevented DJ-1 from translocating to the plasma membrane, thereby inhibited full activation of Lyn, and eventually restrained those receptor-distal signaling molecules, involved Syk, Btk, PLCγ, IP3R, PKC, MAPKs, Akt and NF-κB.
CONCLUSION
Kaempferol could be used as a DJ-1 modulator for preventing MC-mediated allergic disorders through attenuating Lyn activation.
背景
免疫球蛋白 E（IgE）介导的肥大细胞（MC）激活在多种过敏性疾病中至关重要。帕金森病蛋白 7（DJ-1）和 Lyn 激酶被报道为人类 IgE 受体（FcεRI）信号中的受体近端事件。山奈酚是一种主要来源于传统中药莪术根茎的天然类黄酮（姜科），已被证实能抑制过敏反应，但仅限于大鼠嗜碱性白血病细胞的受体远端信号。山奈酚对人类 MC 的抑制作用尚未进行全面研究。
目的
研究山奈酚对体内和体外 IgE 介导的过敏反应的抑制作用及其作用机制，特别是受体近端信号。
方法
应用 IgE 介导的被动皮肤过敏反应和全身过敏反应模型，阐明山奈酚的抗过敏活性。脱颗粒试验、钙成像、细胞因子和趋化因子释放实验用于评估山奈酚在体外的抗过敏作用。采用 Western blot 分析方法研究 DJ-1/Lyn 信号通路及其下游分子。激酶活性测定、免疫荧光和分子对接实验用于证实山奈酚对 DJ-1/Lyn 分子的影响。
结果
山奈酚呈剂量依赖性抑制卵清蛋白/IgE 诱导的小鼠爪肿胀、原发性 MC 从爪皮激活，以及恢复体温过低，降低血清中组胺、肿瘤坏死因子-α、白细胞介素-8 和单核细胞趋化蛋白-1 的浓度。此外，山奈酚抑制 IgE 介导的 LAD2 细胞脱颗粒和钙波动。值得注意的是，山奈酚与 DJ-1 蛋白结合，最初阻止 DJ-1 向质膜移位，从而抑制 Lyn 的完全激活，最终抑制那些受体远端信号分子，包括 Syk、Btk、PLCγ、IP3R、PKC、MAPKs、Akt 和 NF-κB。
结论
山奈酚可作为 DJ-1 调节剂，通过抑制 Lyn 激活来预防 MC 介导的过敏疾病。
Zotero 插件