薤白超细粉通过抑制铁死亡并调控NF-κB/STAT3信号轴缓解脂多糖诱导的小鼠急性肺损伤OA
Ultrafine Xiebai powder alleviates lipopolysaccharide-induced acute lung injury in mice by suppressing ferroptosis and modulating the NF-κB/STAT3 signaling axis
目的 探讨薤白超细粉(UXP)对脂多糖(LPS)诱导急性肺损伤的保护作用及其机制.方法 选取54只雄性C57BL/6小鼠,随机分为对照组、模型组、UXP低剂量组、UXP中剂量组、UXP高剂量组、薤白普通研磨粉(GXP)低剂量组、GXP中剂量组、GXP高剂量组和地塞米松组(LPS+DEX),6只/组.模型组及各干预组小鼠经气管滴注LPS(4 mg/kg)建立急性肺损伤模型.UXP及GXP处理组自造模当日起连续6 d灌胃UXP悬液和薤白普通研磨粉[100、250、500 mg/(kg·d)];地塞米松组灌胃地塞米松溶液[1 mg/(kg·d)];对照组和模型组同期灌胃等体积生理盐水.实验结束后,测定肺组织湿/干重比(W/D)评估肺水肿;采用ELISA检测支气管肺泡灌洗液(BALF)中α肿瘤坏死因子(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素-6(IL-6)及白细胞介素-8(IL-8)水平;流式细胞术分析BALF中炎症细胞数量;HE染色观察肺组织病理学变化;使用试剂盒测定肺组织超氧化物歧化酶(SOD)、丙二醛及谷胱甘肽(GSH)含量;RT-qPCR检测肺组织中TNF-α、IL-1β、IL-6、IL-8及MCP-1 mRNA表达;Western blotting检测p-NF-κB/NF-κB及p-STAT3/STAT3蛋白水平;透射电子显微镜观察线粒体超微结构.细胞实验采用LPS(10 μg/mL)诱导人支气管上皮细胞(Beas-2b)损伤模型,设对照组、模型组、UXP组(LPS+UXP,5 mg/mL)和地塞米松组(LPS+DEX,5 μmol/L).通过CCK-8法测定细胞活性,Annexin V/PI流式检测细胞凋亡,DCFDA探针检测活性氧水平,并利用Mitotracker、Lipofluo和FerroOrange荧光探针评估线粒体膜电位、脂质过氧化及亚铁离子水平.结果 与对照组相比,模型组小鼠W/D比升高(P<0.001),BALF中炎症因子水平上调(P<0.001),炎症细胞数量明显增加,肺组织出现广泛炎症浸润和结构破坏.与此同时,SOD及GSH含量下降(P<0.001),丙二醛水平升高(P<0.001),炎症因子mRNA表达及p-NF-κB/NF-κB、p-STAT3/STAT3蛋白水平增加(P<0.001),线粒体出现体积缩小、嵴减少及膜结构破坏等超微结构损伤.与模型组相比,UXP改善上述指标(P<0.001),部分效果接近地塞米松组.细胞实验结果显示,UXP提高细胞活性(P<0.001),降低凋亡比例(P<0.001),减少活性氧生成,维持线粒体功能,并有效抑制脂质过氧化和铁死亡(P<0.001).结论 UXP对LPS诱导的急性肺损伤具有显著保护作用,其机制可能与抑制炎症因子释放、缓解氧化应激、抑制铁死亡及细胞凋亡、调控NF-κB/STAT3信号通路,以及维持线粒体结构和功能完整性密切相关.
Objective To investigate the protective effects of ultrafine Xiebai powder(UXP)against lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice and the underlying mechanism.Methods Fifty-four male C57BL/6 mice were randomized into 9 groups,including a control group and 8 intratracheal LPS instillation-induced ALI model groups gavaged with saline,dexamethasone(DEX,1 mg/kg),or 100,250,or 500 mg/kg UXP or granule Xiebai powder(n=6)for 6 consecutive days,starting on the day of modeling.After the treatment,the lung wet-to-dry(W/D)ratios were determined,inflammatory cytokines in bronchoalveolar lavage fluid(BALF)were measured by ELISA,and inflammatory cell infiltration was assessed by flow cytometry.Histopathological injury of the lungs was observed using HE staining,and pulmonary SOD,MDA,and GSH levels were determined.Pulmonary mRNA expressions of inflammatory mediators were quantified by RT-qPCR,and NF-κB and STAT3 activation was analyzed using Western blotting;mitochondrial ultrastructure was examined with transmission electron microscopy.In a LPS-stimulated BEAS-2B cell model,the effects of UXP and DEX were assessed on cell viability,apoptosis,reactive oxygen species(ROS),mitochondrial function,lipid peroxidation,and ferrous iron levels.Results The LPS-challenged mice exhibited a higher lung W/D ratio and increased BALF cytokine levels and inflammatory cell counts with pronounced lung inflammation and tissue damage,lowered SOD and GSH levels,elevated MDA levels,increased mRNA expressions of inflammatory mediators and the p-NF-κB/NF-κB and p-STAT3/STAT3 ratios,and obvious mitochondrial damages.UXP markedly alleviated these changes with an efficacy comparable to DEX.In LPS-stimulated BEAS-2B cells,UXP significantly improved cell viability,redcued cell apoptosis and ROS accumulation,preserved mitochondrial integrity,and reduced lipid peroxidation and ferroptosis-associated changes.Conclusion UXP has strong protective effects against LPS-induced ALI in mice possibly by suppressing inflammation,oxidative stress,ferroptosis,and apoptosis,modulating the NF-κB/STAT3 signaling axis,and maintaining mitochondrial integrity.
祝金超;邵宁宁;黄泽彧;刘燕青;孟祥艳;刘子泉;孙予杰;董津睿;樊毫军
天津大学卫生应急学院,医学救援关键技术装备应急管理部重点实验室,天津 300072天津大学医学院,天津 300072天津大学医学院,天津 300072天津大学卫生应急学院,医学救援关键技术装备应急管理部重点实验室,天津 300072天津大学卫生应急学院,医学救援关键技术装备应急管理部重点实验室,天津 300072天津大学卫生应急学院,医学救援关键技术装备应急管理部重点实验室,天津 300072海南万代兰生物医药科技有限公司,海南 澄迈县 571900天津大学医学院,天津 300072天津大学卫生应急学院,医学救援关键技术装备应急管理部重点实验室,天津 300072
薤白超细粉急性肺损伤炎症氧化应激铁死亡
ultrafine Xiebai powderacute lung injuryinflammationoxidative stressferroptosis
《南方医科大学学报》 2026 (5)
1006-1017,12
国家科技部重点研发计划项目(2024YFC3016604)国家应急管理部重点实验室自主创新基金项目(YJBZZJJTJU202403)国家自然科学基金(82200655)国家级大学生创新训练计划项目(202310056123)Supported by National Natural Science Foundation of China(82200655).
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