背景:Calycosin,从黄芪中提取的黄酮类化合物,在室内尘螨诱发的哮喘中显示出抗哮喘的益处。最近的研究表明,先天型细胞,包括第2组先天淋巴细胞(ILC2s)和巨噬细胞,作为2型免疫的诱因和哮喘药物开发的目标。这项工作的重点是calycosin对过敏性哮喘中异常调节的ILC2s和巨噬细胞的影响。
方法:体内,用卵清蛋白(OVA)致敏和攻击建立哮喘小鼠模型,并以20和40mg/kg的剂量腹膜内给药。在体内,用白细胞介素(IL)-33刺激小鼠原代ILC2s,用IL-4和IL-13刺激小鼠RAW264.7巨噬细胞以建立细胞模型。用5μM和10μM剂量的毛蒜素处理细胞。
结果:体内,我们观察到嗜酸性粒细胞数量显著减少,中性粒细胞,暴露于OVA的小鼠支气管肺泡灌洗液(BALF)中的单核细胞巨噬细胞和淋巴细胞。组织病理学评估显示,calycosin抑制OVA引起的气道炎症和重塑。Calycosin显着降低OVA诱导的哮喘小鼠BALF和/或肺组织中上调的IL-4,IL-5,IL-13,IL-33和抑制致瘤性2(ST2)。毛蒜素抑制精氨酸酶1(ARG1)的增加,哮喘小鼠肺组织中IL-10,几丁质酶样3(YM1)和甘露糖受体C型1(MRC1)水平。在体内,calycosin抑制IL-33诱导的活化以及IL-4,IL-5,IL-13和ST2的增加。在RAW264.7巨噬细胞中,毛黄蛋白酶还抑制由IL-4和IL-13诱导的ARG1、IL-10、YM1和MRC1的增加。此外,我们发现,这些变化在40mg/kgcalycosin治疗中比20mg/kgcalycosin治疗更显著。
结论:总的来说,这项研究表明,calycosin可能通过阻止ILC2激活和巨噬细胞M2极化来减轻OVA诱导的哮喘小鼠气道炎症和重塑。我们的研究可能有助于哮喘治疗的进一步研究。
BACKGROUND: Calycosin, a flavonoid compound extracted from Astragalus membranaceus, has shown anti-asthma benefits in house dust mite-induced asthma. Recent studies have suggested that innate-type cells, including group 2 innate lymphoid cells (ILC2s) and macrophages, serve as incentives for type 2 immunity and targets for drug development in asthma. This work focuses on the effects of
calycosin on the dysregulated ILC2s and macrophages in allergic asthma.
METHODS: In vivo, the asthmatic mouse model was established with ovalbumin (OVA) sensitization and challenge, and calycosin was intraperitoneally administered at doses of 20 and 40 mg/kg. In vivo, mouse primary ILC2s were stimulated with interleukin (IL)-33 and mouse RAW264.7 macrophages were stimulated with IL-4 and IL-13 to establish the cell models. Cells were treated with
calycosin at doses of 5 and 10 µM.
RESULTS: In vivo, we observed significantly reduced numbers of eosinophils, neutrophils, monocyte macrophages and lymphocytes in the bronchoalveolar lavage fluid (BALF) of OVA-exposed mice with 40 mg/kg
calycosin. Histopathological assessment showed that calycosin inhibited the airway inflammation and remodeling caused by OVA.
Calycosin markedly decreased the up-regulated IL-4, IL-5, IL-13, IL-33, and suppression tumorigenicity 2 (ST2) induced by OVA in BALF and/or lung tissues of asthmatic mice. Calycosin repressed the augment of arginase 1 (ARG1), IL-10, chitinase-like 3 (YM1) and mannose receptor C-type 1 (MRC1) levels in the lung tissues of asthmatic mice. In vivo, calycosin inhibited the IL-33-induced activation as well as the increase of IL-4, IL-5, IL-13 and ST2 in ILC2s. Calycosin also repressed the increase of ARG1, IL-10, YM1 and MRC1 induced by IL-4 and IL-13 in RAW264.7 macrophages. In addition, we found that these changes were more significant in 40 mg/kg calycosin treatment than 20 mg/kg
calycosin.
CONCLUSIONS: Collectively, this study showed that calycosin might attenuate OVA-induced airway inflammation and remodeling in asthmatic mice via preventing ILC2 activation and macrophage M2 polarization. Our study might contribute to further study of asthmatic therapy.