蛞蝓黄酮通过巨噬细胞对COPD调控的分子机制研究

Chronic obstructive pulmonary disease (COPD) is a major chronic disease characterized by chronic inflammation, and the control of abnormal innate immune activation has become a hotspot in the study of COPD progression. Limax, according to compendium of materia medica records, can be used to treat wheezing, pharyngitis, carbuncle, and so on, we first found that the water extract of Limax has good effect on improving pulmonary inflammation, by preventing mucins overexpression and macrophage invasion in the airway of tobacco smoke-induced COPD mice. According to the mass spectrum analysis and in silico analysis, the natural flavonoid products of Limax have the characteristics of binding with macrophage surface receptor Siglec-1, but what mechanism are these natural products involved in regulating innate immune abnormalities is not clear. Research at home and abroad suggest that mucins can regulate immunity by Siglec receptor and participate in the activation of pyroptosis, hence we proposed that the natural flavonoid products of the Limax inhibit the pyroptosis of macrophages by targeting the Siglec-1 is the key to regulate the abnormal activation of innate immunity. In this project, the COPD mice model and Siglec-1 conditional knockout mice, immunofluorescence and immunoprecipitation, and so on, were designed to obtain the reliable evidence for the inhibition of macrophages abnormal activation by Limax natural flavonoid products. Aim to provide more adequate scientific evidence for the new COPD treating targets development and for considering Limax as candidate drugs for COPD.

慢阻肺(COPD)是一种以慢性炎症为特征的重大慢性疾病，控制固有免疫异常激活已成为遏制COPD进程的研究热点。中药蛞蝓据《本草纲目》等记载可用于治疗喘息、喉痹、痈肿等，我们首次报道蛞蝓水提物能通过减少气道粘蛋白表达、减少巨噬细胞侵润等改善COPD小鼠气道炎症。前期通过代谢谱分析及计算机模拟小分子与受体拟合结果，我们发现蛞蝓黄酮具有靶向巨噬细胞Siglec-1受体特征，但其参与调控固有免疫异常激活的机制尚不清楚。国内外研究提示粘蛋白能通过Siglec受体调节免疫并参与激活细胞焦亡，因此我们提出蛞蝓黄酮通过靶向抑制Siglec-1阻止巨噬细胞焦亡可能是其调控固有免疫异常激活的关键。本项目拟应用小鼠COPD模型和Siglec-1基因敲除小鼠，通过免疫荧光、免疫共沉淀等方法，获得蛞蝓黄酮调控巨噬细胞异常激活的可靠证据，为确立COPD治疗新靶点及将蛞蝓黄酮列为COPD防治候选药物提供充分的科学依据。

慢阻肺(COPD)是一种以慢性炎症为特征的重大慢性疾病，控制固有免疫异常激活已成为遏制COPD进程的研究热点。前期我们发现中药蛞蝓水提物对COPD小鼠气道炎症、肺气肿等损伤有较好的改善，并通过代谢谱分析及计算机模拟小分子与受体拟合结果发现蛞蝓黄酮具有靶向巨噬细胞Siglec-1受体特征，但其参与调控固有免疫异常激活的机制尚不清楚。本项目主要围绕蛞蝓黄酮如何调控固有免疫异常激活而展开。本项目应用小鼠COPD模型和AHR腺病毒过表达细胞株等，对两种蛞蝓黄酮进行了药效学评价。结果表明蛞蝓黄酮F优于黄酮O而更有利于治疗COPD，通过RNA-seq及蛋白质组测序找到其干预前后的差异蛋白并采用WB、QPCR等验证确认其中作用靶点可能是AHR、CYP1A1和AAT，进一步通过小分子-蛋白互作及SPR等手段确认黄酮F与上述关键蛋白的互作关系。本项目为蛞蝓黄酮F调控固有免疫细胞中的中性粒细胞异常激活提供了可靠证据，为确立COPD治疗新靶点及将蛞蝓黄酮列为COPD防治候选药物提供充分的科学依据。