背景:过敏性疾病的发病率不断增加,但其发病机制尚不完全清楚。麻风(SD),在中国被称为“方峰”,不仅可以用于退烧药,镇痛和抗炎作为传统的中药,但也作为活性成分在约8%的处方。然而,其对I型变态反应和假性变态反应的影响尚未明确.
目的:探讨SD及其主要生物活性成分Prim-O-葡萄糖基西米富净(POG)对I型变态反应和假性变态反应的体内外治疗作用及潜在机制。
方法:采用体外RBL-2H3细胞模型和体内被动皮肤过敏(PCA)小鼠模型,评价SD汤和POG对I型变态反应的抑制作用及其可能机制。研究了DNP-IgE/DNP-BSA和化合物48/80(C48/80)诱导的RBL-2H3细胞的细胞脱颗粒,并通过qRT-PCR和WesternBlot分析进一步检测脱颗粒相关信号通路分子。同时,采用PCA模型评价SD汤和POG的体内治疗效果。进行了分子对接技术以探索潜在的机制。
结果:在DNP-IgE/DNP-BSA诱导的细胞模型中,高剂量SD组和POG组β-Hex释放率分别为43.79%和57.01%,高剂量SD组和POG组HA释放量分别为26.19ng/mL和24.20ng/mL。显著低于模型组。此外,SD汤和POG均能明显抑制细胞内Ca2+的增加和细胞凋亡。但对C48/80诱导的细胞脱颗粒无明显影响。分子对接结果表明,5-O-甲基维香苷与POG能与FcεRIα结合,结合能力更强,但对Mrgprx2的结合能力弱。此外,qPCR和Westernblot分析表明SD可以下调Lyn/Syk/PLCγ,MAPK和PI3K/AKT/NF-κB信号通路抑制IgE依赖性细胞脱颗粒。在小鼠PCA模型中,SD和POG都可以剂量依赖性地减弱伊文思蓝外渗,DNP-IgE/DNP-BSA诱导的爪和耳肿胀,但在C48/80诱导的PCA模型下没有明显的抑制作用。
结论:结论:SD对治疗I型过敏有效,表明SD是治疗I型变态反应的潜在候选者,这些影响的潜在机制需要进一步研究。
BACKGROUND: The incidence of allergic disease is constantly increasing, but its pathogenesis is not fully understood. Saposhnikovia divaricata (SD), called \'Fangfeng\' in China, not only can be used for antipyretic, analgesic and anti-inflammatory as a traditional Chinese medicine, but also as an active ingredient in about 8% prescriptions. However, its effects on type I allergy and pseudoallergy have not been clarified.
OBJECTIVE: To explore the treatment and potential mechanisms of SD and its major bioactive component Prim-O-glucosylcimifugin (POG) on type I allergy and pseudoallergy in vitro and in vivo.
METHODS: The inhibitory effect of SD decoction and POG on type I allergy and its possible mechanism were evaluated by using RBL-2H3 cells model in vitro and the passive cutaneous anaphylaxis (PCA) mouse model in vivo. The cell degranulation of RBL-2H3 cells induced by DNP-IgE/DNP-BSA and Compound 48/80 (C48/80) was investigated, and the molecules of degranulation related signaling pathway was further detected by qRT-PCR and Western Blot analysis. Meanwhile, therapeutic effect of SD Decoction and POG were evaluated using PCA models in vivo. The molecular docking technology was conducted to explore the potential mechanisms.
RESULTS: In cells model induced by DNP-IgE/DNP-BSA, the release rate of β-Hex in high dose of SD and POG groups were 43.79% and 57.01%, and the release amount of HA in high dose of SD and POG groups were 26.19 ng/mL and 24.20 ng/mL. They were significantly lower than that in the model group. Besides, SD decoction and POG could significantly inhibit intracellular Ca2+ increasing and cell apoptosis. But there is no obvious effect on cells degranulation induced by C48/80. The molecular docking results showed that 5-O-Methylvisamioside and POG could bind with FcεRI α with stronger binding ability, but weak binding ability to Mrgprx2. Moreover, qPCR and Western blot analyses indicated that SD could down-regulate Lyn/Syk/PLCγ, MAPK and PI3K/AKT/NF-κB signal pathway to inhibit IgE-dependent cell degranulation. In mice PCA model, both SD and POG could dose-dependently attenuate the Evans Blue extravasation, paw and ear swelling induced by DNP-IgE/DNP-BSA, but no significant inhibition under the PCA models induced by C48/80.
CONCLUSIONS: In conclusion, SD is effective for the therapeutic of type I allergies, suggesting that SD is a potential candidate for the treatment of type I allergy, and the underlying mechanism of these effects needs to be further studied.