蓝萼甲素通过抑制中性粒细胞NETs的形成缓解急性肺损伤-现代医学

蓝萼甲素通过抑制中性粒细胞NETs的形成缓解急性肺损伤

单位：1. 苏州大学附属第一医院江苏省血液研究所, 国家血液系统疾病临床医学研究中心, 国家卫生健康委血栓与止血重点实验室, 血液学协同创新中心, 江苏苏州 215006;
2. 苏州大学放射医学与辐射防护国家重点实验室, 江苏苏州 215006

关键词：蓝萼甲素急性肺损伤中性粒细胞中性粒细胞胞外陷阱

分类号：R285.5

出版年·卷·期（页码）：2025·44·第六期（869-876）

摘要：

目的： 研究蓝萼甲素(GLA)对ALI的治疗作用及可能机制，为临床治疗提供理论依据。方法： 将C57小鼠分为正常对照组、脂多糖(LPS)组、GLA组、LPS+GLA低剂量(5 mg·kg^-1)组、LPS+GLA高剂量(10 mg·kg^-1)组和LPS+地塞米松(DEX)(5 mg·kg^-1)组，每组8~10只小鼠。通过鼻腔滴注将LPS注入气管建立小鼠ALI模型。于建模前2 h和建模后2 h腹腔注射GLA。建模24 h后，抽取肺泡灌洗液(BALF)进行各项指标检测。取左肺组织称重，计算干重/湿重；HE染色观察肺组织病理变化；酶联免疫吸附试验(ELISA)检测BALF中白介素(IL)-6、IL-1β、肿瘤坏死因子-α(TNF-α)含量；免疫荧光染色观察肺组织中性粒细胞聚集及中性粒细胞细胞外陷阱(NETs)的生成。结果： 与正常对照组相比，LPS组出现肺泡塌陷、肺泡壁厚度增加等病理现象，引发严重的肺部损伤，中性粒细胞增多、肺部炎症因子水平升高。相比之下，GLA组和DEX组肺部损伤减轻，并且肺部出血没有增加。同时GLA组肺组织中性粒细胞活化减少，这与NETs形成减少有关，GLA抑制NETs的形成，并显著抑制肽基精氨酸脱氨酶4(PAD4)蛋白。结论： GLA可能通过抑制肺部中性粒细胞NETs的形成来缓解小鼠ALI的发病。

Objective: This study aimed to explore the therapeutic role of GLA in ALI and its underlying mechanisms, providing a theoretical basis for clinical treatment. Methods: C57 mice were divided into the followinggroups(n=8-10): normal control group, LPS group, GLA group, LPS+low-dose GLA(5 mg·kg^-1) group, LPS+high-dose(10 mg·kg^-1) GLA group, and LPS+dexamethasone(DEX, 5 mg·kg^-1) group. The ALI model was established by intranasal instillation of LPS. GLA was intraperitoneally administered 2 hours before and 2 hours after LPS exposure. At 24 hours post-modeling, bronchoalveolar lavage fluid(BALF) and lung tissues were collected. Lung injury severity was evaluated by wet-to-dry weight ratio and hematoxylin-eosin(HE) staining. ELISA was used to measure interleukin-6(IL-6), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α) levels in BALF. Neutrophil infiltration and neutrophil extracellular traps(NETs) in lung tissue were analyzed by immunofluorescence staining.Results: Compared with the normal control group, the LPS group demonstrated severe lung injury with alveolar collapse, thickened alveolar walls, increased neutrophil infiltration, and elevated inflammatory cytokines(IL-6, IL-1β, TNF-α). Both GLA and DEX groups exhibited alleviated lung damage without aggravated pulmonary hemorrhage. GLA treatment significantly reduced neutrophil activation in lung tissue, which was closely associated with marked inhibition of NETs formation. Furthermore, GLA downregulated the expression of peptidyl arginine deiminase 4(PAD4) protein. Conclusion: GLA alleviates ALI in mice by suppressing the formation of NETs in lung tissues.

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