玉屏风散通过ZBP1介导的巨噬细胞泛凋亡抑制哮喘气道炎症的机制研究  

Mechanism of Yupingfeng powder inhibiting airway inflammation in asthma through ZBP1⁃mediated alveolar macrophage PANoptosis

作　　者：安琪 李利清[1,2] 刘晓 虞坚尔[1,2] AN Qi;LI Liqing;LIU Xiao;YU Jianer(Department of Pediatrics,Shanghai Traditional Chinese Medicine Hospital Affiliated to Shanghai University of Chinese Medicine,Shanghai 200071,China;Institute of Pediatrics of Traditional Chinese Medicine,Shanghai Academy of Traditional Chinese Medicine,Shanghai 200071,China)

机构地区：[1]上海中医药大学附属上海市中医医院儿科,上海200071 [2]上海市中医药研究院中医儿科研究所,上海200071

出　　处：《海南医学院学报》2024年第12期905-912,共8页Journal of Hainan Medical University

基　　金：第五批全国中医临床优秀人才研修项目[国中医药人教函(2022)1号]。

摘　　要：目的:观察玉屏风散对ZBP1介导的肺泡巨噬细胞泛凋亡的影响,从而阐明其抑制哮喘气道炎症的分子机制。方法:采用脂多糖诱导肺泡巨噬细胞(MH‑S)构建哮喘细胞模型,并给予不同浓度玉屏风散含药血清进行干预,分别采用CCK8法检测NH‑S细胞活性,ELISA法检测LDH、IL‑1β和IL‑18的含量,流式细胞术检测细胞死亡情况,Western blot法检测泛凋亡相关蛋白和ZBP1蛋白的表达,RT‑qPCR法检测ZBP1基因的表达。结果:与空白组相比,模型组肺泡巨噬细胞LDH释放、IL‑1β与IL‑18的含量和死亡细胞比例显著增加,细胞焦亡蛋白Cleaved caspase‑1与GSDMD‑NT、坏死性凋亡蛋白p‑RIPK3与p‑MLKL、凋亡蛋白Cleaved caspase‑3的表达显著上调,差异均具有统计学意义(P<0.01)。与模型组相比,玉屏风散组肺泡巨噬细胞LDH释放、IL‑1β与IL‑18的含量和死亡细胞比例显著降低,细胞焦亡蛋白Cleaved caspase‑1与GSDMD‑NT、坏死性凋亡蛋白p‑RIPK3与p‑MLKL、凋亡蛋白Cleaved caspase‑3的表达显著下调,差异均具有统计学意义(P<0.01)。此外,与模型组相比,玉屏风散组肺泡巨噬细胞中ZBP1蛋白和ZBP1mRNA的表达显著降低,差异均具有统计学意义(P<0.01)。结论:玉屏风散通过抑制巨噬细胞中ZBP1的表达,从而抑制细胞泛凋亡水平改善气道炎症。Objective:To observe the effect of Yupingfeng powder on ZBP1‑mediated PANoptosis of alveolar macrophages so as to elucidate its molecular mechanism of inhibiting airway inflammation in asthma.Methods:Lipopolysaccharide was used to induce alveolar macrophages(MH‑S)to construct an asthma cell model,and different concentrations of Yupingfeng powder medicated serum were given for intervention.CCK8 method was used to detect the activity of NH‑S cells,ELISA method was used to detect the content of LDH,IL‑1βand IL‑18,flow cytometry was used to detect cell death,Western blot method was used to detect the expression of PANoptosis‑related proteins and ZBP1 protein,and RT‑qPCR method was used to detect the expression of ZBP1 gene.Results:Compared with the control group,the LDH release of alveolar macrophages,the content of IL-1βand IL-18 and the proportion of dead cells in the model group were significantly increased.The expressions of pyroptosis proteins(Cleaved caspase-1 and GSDMD-NT),necroptosis proteins(p-RIPK3 and p-MLKL),and apoptosis protein(Cleaved caspase-3)were significantly up-regulated(P<0.01).Compared with the model group,the expression of LDH release,the content of IL-1βand IL-18,and the proportion of dead cells in alveolar macrophages in the Yupingfeng powder group were significantly reduced,and the expression of Cleaved caspase-1,GSDMD-NT,p-RIPK3,p-MLKL,and Cleaved caspase-3 were significantly down-regulated(P<0.01).In addition,the expression of ZBP1 protein and ZBP1 mRNA in alveolar macrophages in the Yuping-feng powder group was significantly reduced compared with the model group,and the differences were statistically significant(P<0.01).Conclusion:Yupingfeng powder ameliorates airway inflammation by inhibiting the expression of ZBP1 in alveolar macro-phages,thereby suppressing the level of PANoptosis.

关 键 词：玉屏风散 肺泡巨噬细胞 泛凋亡 哮喘 气道炎症 

分 类 号：R285[医药卫生—中药学]