南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (2): 381-386.doi: 10.12122/j.issn.1673-4254.2024.02.21

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茶多酚通过抑制NLRP3炎症小体改善脓毒症小鼠的急性肺损伤

凌旭光，徐雯雯，庞观来，洪旭星，刘凤芹，李 洋   

  1. 南方医科大学南方医院健康管理科，门诊部，广东 广州 510515；南方医科大学珠江医院消化内科，广东 广州 510280
  • 发布日期:2024-03-14

Tea polyphenols ameliorates acute lung injury in septic mice by inhibiting NLRP3 inflammasomes

LING Xuguang, XU Wenwen, PANG Guanlai, HONG Xuxing, LIU Fengqin, LI Yang   

  1. Department of Health Management, Outpatient Department, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Department of Gastroenterology, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China
  • Published:2024-03-14

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摘要： 目的 探讨茶多酚（TP）对NLRP3炎症小体的调控作用机制以及其对脓毒症相关急性肺损伤的治疗效用。方法 将60只C57BL/6小鼠随机平均分为假手术组（单纯开腹探查盲肠后即关腹）、盲肠结扎穿孔组（开腹暴露并游离盲肠后行盲肠结扎与穿孔操作后关腹）以及盲肠结扎穿孔+TP治疗组（盲肠结扎穿孔前1周予TP灌胃），每组20只。建模完成后绘制生存曲线评估各组小鼠对脓毒症打击的耐受情况。测定各组小鼠肺湿/干质量比值、肺组织髓过氧化物酶（MPO）水平，并对肺组织进行HE染色和急性肺损伤评分，评估肺损伤程度。ELISA法测定肺组织内IL-1β、TNF-α及IL-6表达水平；蛋白免疫印迹实验检测肺组织内NLRP3、caspase-1 p10及ASC蛋白表达水平；免疫组化染色测定肺组织内MPO、caspase-8及NLRP3蛋白表达，评估各组小鼠肺内炎症情况。检测肺组织内MDA及H2O2以评估氧化应激水平；免疫荧光共染NLRP3和NOX4蛋白以探讨二者表达及共定位。结果 TP治疗组小鼠术后72 h死亡率、小鼠肺湿/干质量比值、肺组织MPO水平以及急性肺损伤评分均较盲肠结扎穿孔组小鼠显著下降（P<0.05）；TP治疗组小鼠肺组织内IL-1β、TNF-α及IL-6、NLRP3、caspase-1 p10、ASC等蛋白表达水平亦较盲肠结扎穿孔组小鼠显著降低（P<0.05）；氧化应激相关检测结果提示，TP治疗组小鼠肺组织内MDA及H2O2水平较盲肠结扎穿孔组小鼠显著减低（P<0.05）；免疫荧光共染则提示TP治疗组小鼠NOX4蛋白与NLRP3蛋白在肺组织内的表达和共定位较盲肠结扎穿孔组小鼠减少。结论 TP可通过抑制NLRP3炎症小体相关炎症改善盲肠结扎穿孔诱导的小鼠脓毒症肺损伤，且这一调控机制可能与其对肺组织内氧化应激相关蛋白NOX4的表达抑制相关。

关键词: 茶多酚；脓毒症；急性肺损伤；NLRP3炎症小体；氧化应激

Abstract: Objective To investigate the mechanism of tea polyphenols (TP) for regulating NLRP3 inflammasomes and alleviating acute lung injury in septic mice. Methods Sixty C57BL/6 mice were randomly assigned into sham-operated, cecal ligation and puncture (CLP) and CLP + TP treatment groups, and survival of the mice was recorded after modeling in each group. The lung wet/dry weight ratio and myeloperoxidase (MPO) activity were determined, and lung injury of the mice was evaluated using HE staining and acute lung injury score. The expressions of IL-1β, TNF-α, IL-6, NLRP3, caspase-1 p10, ASC, MPO, and caspase-8 in the lung tissue were detected using ELISA, Western blotting, or immunohistochemical staining. MDA and H2O2 levels in the lungs were detected to evaluate the level of oxidative stress. Immunofluorescence assay was used to investigate the co-localization of NLRP3 and NOX4. Results The postoperative mortality rate at 72 h, lung wet/dry weight ratio, MPO level and acute lung injury scores were significantly lower in CLP+TP group than in CLP group (P<0.05). Treatment with TP significantly reduced the expressions of NLRP3- related inflammatory factors (P<0.05) and lowered MDA and H2O2 levels in the lung tissue of the septic mice (P<0.05). Immunofluorescence co-staining showed a lower level of NOX4 and NLRP3 co-localization in CLP+TP group than in CLP group. Conclusion TP inhibits NLRP3 inflammasome-associated inflammation to alleviate CLP-induced acute lung injury in mice through a regulatory mechanism that inhibits NOX4 expression and reduces oxidative stress in the lung tissue.

Key words: tea polyphenols; sepsis; acute lung injury; NLRP3 inflammasomes; oxidative stress