甘草黄酮B通过抑制上皮PI3K-AKT/NF-κB NLRP3炎症小体信号减轻过敏性鼻炎

文献信息

中文摘要

过敏性鼻炎仍然高度流行，且现有治疗往往控制不完全，同时越来越多的证据提示上皮PI3K-AKT/NF-κB介导的NLRP3炎症小体激活参与上气道炎症。本研究探讨甘草来源的黄酮类化合物甘草黄酮B（LFB）是否通过调节该信号轴减轻过敏性鼻炎。在卵清蛋白诱导的小鼠模型中，评估过敏性鼻炎样症状和鼻部组织病理学改变，采用ELISA定量细胞因子水平，并通过免疫印迹评估NLRP3炎症小体及PI3K-AKT/NF-κB信号的激活。随后进行无偏RNA测序及KEGG和GO富集分析，以识别潜在的上游通路。通过体内使用IGF-1重新激活PI3K-AKT以及使用poly(I)刺激TLR3，并在原代人鼻上皮细胞（HNEpCs）中进一步探讨PI3K-AKT/NF-κB信号的参与。还进行了分子对接以探索LFB与PI3K之间的潜在相互作用。LFB减少打喷嚏和鼻部抓挠，部分改善上皮增厚和炎症细胞浸润，并减少肥大细胞染色。LFB还降低IL-1β、IL-6、IL-5、IL-13和TNF-α水平，以及NLRP3炎症小体组分的表达。RNA测序分析确定PI3K-AKT信号是主要富集通路之一，免疫印迹证实LFB降低PI3K、AKT和p65的磷酸化，而未显著改变其总蛋白水平。在HNEpCs中，IGF-1恢复通路磷酸化并部分恢复细胞因子和炎症小体蛋白表达，从而减弱LFB的作用。同样，IGF-1或poly(I)在体内部分削弱LFB诱导的症状和组织学改善。分子对接将LFB定位于PI3K催化口袋内，为潜在的LFB-PI3K相互作用提供结构合理性。总之，这些发现表明LFB减轻卵清蛋白诱导的过敏性鼻炎样炎症，并与鼻组织、HNEpCs和刺激的肥大细胞模型中PI3K-AKT/NF-κB/NLRP3信号轴激活减少相关。IGF-1或poly(I)对这些作用的部分逆转进一步支持该信号通路的参与。

英文摘要

Allergic rhinitis remains highly prevalent and is often incompletely controlled by current therapies, while accumulating evidence implicates epithelial PI3K-AKT/NF-κB-mediated activation of the NLRP3 inflammasome in upper-airway inflammation. This study investigated whether the licorice-derived flavonoid Licoflavone B (LFB) alleviates allergic rhinitis by modulating this signaling axis. In an ovalbumin-induced murine model, allergic rhinitis-like symptoms and nasal histopathological changes were evaluated, cytokine levels were quantified by ELISA, and the activation of the NLRP3 inflammasome and PI3K-AKT/NF-κB signaling was assessed by immunoblotting. Unbiased RNA sequencing followed by KEGG and GO enrichment analyses was performed to identify potential upstream pathways. The involvement of PI3K-AKT/NF-κB signaling was further explored through PI3K-AKT reactivation with IGF-1 and TLR3 stimulation with poly(I) in vivo and in primary human nasal epithelial cells (HNEpCs). Molecular docking was also conducted to explore the potential interaction between LFB and PI3K. LFB reduced sneezing and nasal scratching, partially ameliorated epithelial thickening and inflammatory-cell infiltration, and decreased mast-cell staining. LFB also reduced the levels of IL-1β, IL-6, IL-5, IL-13, and TNF-α, as well as the expression of NLRP3-inflammasome components. RNA-sequencing analysis identified PI3K-AKT signaling as one of the major enriched pathways, and immunoblotting confirmed that LFB decreased the phosphorylation of PI3K, AKT, and p65 without substantially altering their total protein levels. In HNEpCs, IGF-1 restored pathway phosphorylation and partially reinstated cytokine and inflammasome protein expression, thereby attenuating the effects of LFB. Similarly, IGF-1 or poly(I) partially diminished the symptomatic and histological improvements induced by LFB in vivo. Molecular docking positioned LFB within the PI3K catalytic pocket, providing structural plausibility for a potential LFB-PI3K interaction. Collectively, these findings indicate that LFB alleviates ovalbumin-induced allergic rhinitis-like inflammation and is associated with reduced activation of the PI3K-AKT/NF-κB/NLRP3 signaling axis in nasal tissues, HNEpCs, and stimulated mast-cell models. The partial reversal of these effects by IGF-1 or poly(I) further supports the involvement of this signaling pathway.