目的 基于核酸结合寡聚结构域（NOD）样受体热蛋白结构域3（NLRP3）炎症小体的激活，探究汉黄芩素对脂多糖（LPS）诱导的小鼠急性肺损伤的治疗作用。方法 C57BL/6J小鼠随机分为对照组、模型组和汉黄芩素低、高剂量（40、80 mg·kg^-1）组，汉黄芩素ip给药3 d，对照组和模型组ip等量0.9%氯化钠溶液。第3次给药1 h后给予模型组和汉黄芩素低、高剂量组小鼠ip 25 mg·kg^-1的LPS，对照组ip等量0.9%氯化钠溶液。统计小鼠死亡情况，绘制生存曲线； LPS刺激12 h后监测小鼠呼吸频率；苏木精-伊红（HE）染色检测肺组织病理变化；实时荧光定量PCR（qRT-PCR）检测肺组织炎症相关白细胞介素-1β（Il1b）、肿瘤坏死因子α（Tnfα） mRNA水平； Western blotting检测小鼠肺组织NLRP3、凋亡相关的斑点样蛋白（ASC）、天冬氨酸特异性的半胱氨酸蛋白水解酶-1（cleaved Caspase-1）、Gasdermin D-NT（GSDMD-NT）的蛋白表达。采用200 ng·mL^-1 LPS分别联合4 mmol·L^-1三磷酸腺苷（ATP）、500 mg·mL^-1明矾（Alum）或1 μmol·L^-1短杆菌肽（ Gra）刺激PMs细胞构建经典的NLRP3炎症小体活化模型，Western blotting检测汉黄芩素（5、10、20 μmol·L^-1）对NLRP3炎症小体相关蛋白NLRP3、cleaved Caspase-1、IL-1β和细胞焦亡相关蛋白GSDMD-NT表达的影响。结果 在小鼠急性肺损伤模型中，与模型组比较，汉黄芩素80 mg·kg^-1组小鼠的死亡率显著降低（P＜0.05）；汉黄芩素显著改善小鼠呼吸频率降低（P＜0.05），显著改善小鼠肺组织损伤（P＜0.001），显著降低肺组织Il1b和Tnfα mRNA表达水平（P＜0.001），显著降低肺组织NLRP3、GSDMD-NT蛋白的表达量（P＜0.01、0.001）。在细胞模型中，与模型组比较，汉黄芩素显著降低IL-1β、cleavedCaspase-1、GSDMD-NT蛋白的表达水平（P＜0.05、0.01、0.001）。结论 汉黄芩素通过抑制NLRP3炎症小体活化改善LPS诱导的小鼠急性肺损伤。

Objective To explore the therapeutic effects of wogonin on lipopolysaccharide (LPS) - induced acute lung injury in mice based on the activation of the nucleotide oligomerization domain (NOD) like receptor family pyrin domain-containing 3 (NLRP3) inflammasome. Methods C57BL/6J mice were randomly divided into the control group, model group, and low-dose (40 mg·kg^-1) and high-dose (80 mg·kg^-1) wogonin groups. Wogonin was ip administered for 3 d, while the control and model groups were ip injected with the same volume of 0.9% sodium chloride solution. One hour after the third administration, the model group and the low-dose and high-dose wogonin groups were ip injected with 25 mg·kg^-1 LPS, and the control group was ip injected with the same volume of 0.9% sodium chloride solution. The mortality of mice was recorded, and the survival curve was plotted. The respiratory rate of mice was monitored 12 hours after LPS stimulation. Hematoxylin-eosin (HE) staining was used to detect the pathological changes of lung tissue. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the mRNA levels of inflammation-related interleukin-1β (Il1b) and tumor necrosis factor α (Tnfα) in lung tissue. Western blotting was used to detect the protein expression of NLRP3, apoptosis-associated speck-like protein (ASC), cleaved caspase-1, and gasdermin D-NT (GSDMD-NT) in mouse lung tissue. PMs cells were stimulated with 200 ng·mL^-1 LPS in combination with 4 mmol·L^-1 ATP, 500 mg·mL^-1 Alum, or 1 μmol·L^-1 nigericin to establish a classic NLRP3 inflammasome activation model. Western blotting was used to detect the effects of wogonin (5, 10, and 20 μmol·L^-1) on the expression of NLRP3 inflammasome-related proteins NLRP3, cleaved caspase-1, IL-1β, and pyroptosisrelated protein GSDMD-NT. Results In the acute lung injury model of mice, compared with the model group, the mortality rate of the 80 mg·kg^-1 wogonin group was significantly reduced (P < 0.05); wogonin significantly improved the decreased respiratory rate of mice (P < 0.05), significantly improved lung tissue injury (P < 0.001), significantly reduced the mRNA expression levels of Il1b and Tnfα in lung tissue (P < 0.001), and significantly reduced the protein expression levels of NLRP3 and GSDMD-NT in lung tissue (P < 0.01, 0.001). In the cell model, compared with the model group, wogonin significantly reduced the expression levels of IL-1β, cleaved caspase-1, and GSDMD-NT proteins (P < 0.05, 0.01, 0.001). Conclusion Wogonin ameliorates LPS induced acute lung injury in mice through the inhibition of NLRP3 inflammasome activation.

R285.5

国家自然科学基金面上项目（82474153）；2023年度北京中医药大学-优莎纳联合研究中心（BURC）基金重点项目（BUCM-2022-JS-KF-003）