目的 基于NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor pyrin domain containing 3，NLRP3）/半胱氨酸天冬氨酸蛋白酶-1（cystein-asparate protease-1，Caspase-1）信号通路和p38丝裂原活化蛋白激酶（mitogen-activated protein kinase，MAPK）/c-Jun氨基末端激酶（c-Jun N-terminal kinase，JNK）信号通路探讨清咽滴丸缓解肺部炎症的作用机制。方法 50只雄性Balb/C小鼠随机分为对照组、模型组、地塞米松（5 mg/kg）组和清咽滴丸低、高剂量（0.2、0.8 g/kg）组，给药干预7 d。末次给药1 h后，除对照组外，其余小鼠ip脂多糖（5 mg/kg）诱导急性炎症损伤模型。造模4 h后，取血清和肺组织，计算肺组织湿干质量比；采用ELISA法检测血清中白细胞介素-18（interleukin-18，IL-18）、IL-1β、IL-6、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）水平；苏木素-伊红（hematoxylin-eosin，HE）染色检测肺组织病理学变化；免疫组化法检测肺组织中p-p38 MAPK蛋白表达；Western blotting检测肺组织中p-p38 MAPK、p-JNK、B细胞淋巴瘤-2相关X蛋白（Bcl-2-associated X protein，Bax）、NLRP3、B细胞淋巴瘤-3（B-cell lymphoma-3，Bcl-3）、硫氧还蛋白相互作用蛋白（thioredoxin-interacting protein，TXNIP）和Caspase-1的表达；DHE染色法检测肺组织中活性氧（reactive oxygen species，ROS）水平；TUNEL染色检测肺组织中细胞焦亡水平。结果 与模型组比较，清咽滴丸可显著降低LPS诱导的小鼠肺组织湿干质量比（P＜0.05），改善肺损伤，显著下调血清中IL-18、IL-6、TNF-α和IL-1β水平（P＜0.05、0.01），降低肺组织中p38 MAPK和JNK的磷酸化水平（P＜0.01、0.001），显著下调肺组织中Bax、NLRP3、Bcl-3、TXNIP和Caspase-1蛋白表达（P＜0.001），降低ROS的过度积累（P＜0.01、0.001），抑制细胞焦亡（P＜0.05、0.01、0.001）。结论 清咽滴丸可显著改善LPS诱导的小鼠肺部炎症，其作用机制为通过抑制NLRP3/Caspase-1通路介导的细胞焦亡和p38 MAPK/JNK通路激活协同发挥抗炎作用。

Objective To investigate the mechanism of Qingyan Dropping Pills (清咽滴丸, QDP) in alleviating pulmonary inflammation based on NOD-like receptor pyrin domain containing 3 (NLRP3)/cystein-asparate protease-1 (Caspase-1) and p38 mitogen-activated protein kinase (p38 MAPK)/c-Jun N-terminal kinase (JNK) signaling pathways. Methods A total of 50 male Balb/C mice were randomly divided into control group, model group, dexamethasone (5 mg/kg) group, QDP low-, high-dose (0.2, 0.8 g/kg) groups, and mice were intervened with drugs for 7 d. One hour after the last administration, except for the control group, all other mice were induced with lipopolysaccharide (5 mg/kg) to form an acute inflammatory injury model. After 4 h of modeling, serum and lung tissue were collected to calculate the wet dry weight ratio of lung tissue. Levels of interleukin-18 (IL-18), IL-1β, IL-6 and tumor necrosis factor-α (TNF-α) in serum were measured by ELISA. Histopathological changes in lung were evaluated by hematoxylin-eosin (HE) staining. Expression of p-p38 MAPK was assessed by immunohistochemistry. Western blotting was employed to analyze protein levels of p-p38 MAPK, p-JNK, Bcl-2-associated X protein (Bax), NLRP3, B-cell lymphoma-3 (Bcl-3), thioredoxin-interacting protein (TXNIP) and Caspase-1 in lung tissue. Reactive oxygen species (ROS) level was evaluated by dihydroethidium (DHE) staining. Pyroptosis level in lung tissue was detected by TUNEL staining. Results Compared with model group, QDP could significantly reduce the wet dry weight ratio of lung tissue in LPS-induced mice (P < 0.05), improve lung injury, significantly down-regulate the levels of IL-18, IL-6, TNF-α and IL-1β in serum (P < 0.05, 0.01), reduce the phosphorylation levels of p38 MAPK and JNK in lung tissue (P < 0.01, 0.001), significantly down-regulate the protein expressions of Bax, NLRP3, Bcl-3, TXNIP and Caspase-1 in lung tissue (P < 0.001), reduce the excessive accumulation of ROS (P < 0.01, 0.001), and inhibit cell apoptosis (P < 0.05, 0.01, 0.001). Conclusion QDP could significantly improve LPS-induced lung inflammation in mice, and its mechanism is related to synergistically exert anti-inflammatory effects by inhibiting NLRP3/Caspase-1 pathway mediated cell apoptosis and p38 MAPK/JNK pathway activation.

R285.5

现代中医药海河实验室科技项目（25HHZYSS00006）；国家中医药管理局科技项目（GZY-KJS-2025-063）