Abstract：
Guominkang (GMK), a Chinese medicine formula, has been used to treat allergic diseases in clinical settings for many years. To evaluate the antiallergic effect and molecular mechanism of action of GMK extract, RBL-2H3 cell models and passive cutaneous anaphylaxis (PCA) mouse models were established. High performance liquid chromatography (HPLC) and ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) analyses were performed to characterize the chemical composition of GMK. A total of 94 compounds were identified or tentatively identified from GMK. Three of them, emodin, ursolic acid, and hamaudol, were identified for the first time as potential active compounds in GMK, since they inhibited the degranulation of mast cells. The anti-allergic effect of hamaudol was the first to be discovered. GMK could markedly mitigate the shade of Evans Blue extravasation and ear incrassation in PCA mouse models. Additionally, GMK significantly inhibited the degranulation of mast cells, suppressed mast cell degranulation by reducing Ca2+ influx and the levels of TNF-α, IL-4, and histamine, and markedly inhibited the phosphorylation of Lyn, Syk, PLCγ1, IκBα, and NF-κB p65. Molecular docking results indicated that hamaudol and emodin had strong interaction with FcɛRI and NF-κB related proteins, while ursolic acid only interacted with NF-κB associated proteins. These results suggest GMK suppresses the activation of MCs both in vivo and in vitro. The underlying mechanism of its anti-allergic activity is associated with the inhibition of FcɛRI and NF-κB activation.
摘要：
国明康(GMK)，中药配方，多年来一直被用于临床治疗过敏性疾病。评价GMK提取物的抗过敏作用及其分子机制,建立RBL-2H3细胞模型和被动皮肤过敏反应(PCA)小鼠模型。进行了高效液相色谱（HPLC）和超高效液相色谱-质谱（UHPLC-MS）分析以表征GMK的化学组成。从GMK中鉴定或初步鉴定了总共94个化合物。其中三个,大黄素,熊果酸,和Hamaudol,首次被鉴定为GMK中的潜在活性化合物，因为它们抑制了肥大细胞的脱颗粒。首先发现了hamaudol的抗过敏作用。在PCA小鼠模型中，GMK可以显着减轻伊文思蓝外渗和耳廓的阴影。此外,GMK显著抑制肥大细胞脱颗粒,通过减少Ca2+内流和TNF-α水平抑制肥大细胞脱颗粒，IL-4和组胺,并显著抑制了Lyn的磷酸化，Syk,PLCγ1,IκBα,和NF-κBp65。分子对接结果表明,大黄醇和大黄素与FcºRI和NF-κB相关蛋白有较强的相互作用,而熊果酸仅与NF-κB相关蛋白相互作用。这些结果表明GMK在体内和体外均抑制MC的活化。其抗过敏活性的潜在机制与FcRI和NF-κB激活的抑制有关。
