铁死亡在大鼠烧冲复合伤合并急性肺损伤中的作用及其机制  

Role and mechanism of ferroptosis in combined burn-blast injury with acute lung injury in rats

作　　者：张浩 官浩 汪宇航 张万福 田林强[1] 任文杰 Zhang Hao;Guan Hao;Wang Yuhang;Zhang Wanfu;Tian Linqiang;Ren Wenjie(Henan Medical Key Laboratory for Research of Trauma and Orthopedics,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang453003,China;Department of Burns and Cutaneous Surgery,Burn Center of PLA,the First Affiliated Hospital of Air Force Medical University,Xi'an710032,China)

机构地区：[1]新乡医学院第三附属医院,河南省创伤与骨科医学重点实验室,新乡453003 [2]空军军医大学第一附属医院全军烧伤中心,烧伤与皮肤外科,西安710032

出　　处：《中华烧伤与创面修复杂志》2024年第11期1034-1042,共9页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金面上项目(82272268);新乡医学院河南省创伤骨科重点实验室开放研究基金(HZKFKT20220503)。

摘　　要：目的探讨铁死亡在大鼠烧冲复合伤合并急性肺损伤中的作用及其机制。方法该研究为实验研究。取24只8周龄雄性SD大鼠,按照随机数字表法分为对照组和实验组,每组12只。对实验组大鼠在麻醉后进行爆炸处理以制作烧冲复合伤合并急性肺损伤模型,对照组大鼠行模拟致假伤处理。伤后24 h,采用苏木精-伊红染色和免疫组织化学染色观测肺组织的病理形态,采用酶联免疫吸附测定法检测支气管肺泡灌洗液(BALF)的上清液中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)及IL-6的水平,采用全自动动物血气分析仪检测腹主动脉血的动脉血氧分压(PaO_(2))和动脉血二氧化碳分压(PaCO_(2)),称重肺组织并计算其湿干重比,采用二喹啉甲酸法测定BALF中的总蛋白浓度,基于苏木精-伊红染色对肺损伤情况进行评分,采用相关试剂盒检测大鼠肺组织匀浆液中氧化应激因子活性氧、丙二醛、超氧化物歧化酶(SOD)、谷胱甘肽、亚铁离子水平,采用免疫荧光法和免疫组织化学法检测肺组织中铁死亡相关分子谷胱甘肽过氧化物酶4(GPX4)、脂质过氧化相关分子4-羟基壬烯醛(4-HNE)及DNA氧化损伤相关分子8-羟基脱氧鸟苷(8-OHdG)的表达量,采用透射电子显微镜观察肺组织细胞中线粒体形态。样本数均为6。结果伤后24 h,对照组大鼠肺组织结构清晰完整,肺泡壁正常;实验组大鼠的肺组织水肿明显,肺泡壁变厚、结构不清晰。伤后24 h,与对照组相比,实验组大鼠BALF的上清液中TNF-α、IL-1β及IL-6水平均显著升高(t值分别为3.96、9.84、10.60,P<0.05);实验组大鼠肺组织湿干重比、肺损伤评分以及BALF中总蛋白浓度均明显升高(t值分别为6.91、6.64、10.04,P<0.05),腹主动脉血的PaO_(2)明显下降(t=8.85,P<0.05)而PaCO_(2)未见明显变化(P>0.05);实验组大鼠的肺组织匀浆液中SOD及谷胱甘肽水平均明显降低(t值分别为4.36和8.56,P<0.05),活性氧、�ObjectiveTo investigates the role and mechanism of ferroptosis in combined burn-blast injury with acute lung injury in rats.MethodsThis study was an experimental study.Twenty-four 8-week-old male Sprague-Dawley rats were divided into control group and experimental group by random number table method,each containing 12 animals.The rats in experimental group were anesthetized and subjected to explosion treatment to create the model of combined burn-blast injury with acute lung injury,whereas the rats in control group underwent sham injury.At 24 hours post injury,the pathological morphology of lung tissue was observed by hematoxylin-eosin staining and immunohistochemical staining.The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and IL-6 in the supernatant of bronchoalveolar lavage fluid(BALF)were detected by enzyme-linked immunosorbent assay.The arterial partial pressure of oxygen(PaO_(2))and arterial partial pressure of carbon dioxide(PaCO_(2))of abdominal aortic blood were measured by automatic animal blood gas analyzer.The lung tissue was weighed and the wet-dry weight ratio was calculated.The total protein concentration in BALF was measured by bicinchoninic acid assay.Lung injury was scored based on hematoxylin-eosin staining.The levels of oxidative stress factors,such as reactive oxygen species,malondialdehyde,superoxide dismutase(SOD),glutathione,and ferrous ion in lung tissue homogenate of rats were detected by related kits.The expression levels of ferroptosis-related molecule glutathione peroxidase 4(GPX4),lipid peroxidation-related molecule 4-hydroxynonenal(4-HNE),and oxidative DNA damage-related molecule 8-hydroxydeoxyguanosine(8-OHdG)in lung tissue were detected by immunofluorescence and immunohistochemistry methods.Mitochondrial morphology in lung tissue cells was observed under transmission electron microscopy.The sample number was all 6.ResultsAt 24 hours post injury,the lung tissue structure of rats in control group was clear and complete,and the alveolar wall was normal;in exper

关 键 词：烧伤 吸入性 急性肺损伤 氧化性应激 烧冲复合伤 铁死亡 

分 类 号：R644[医药卫生—外科学]