摘要
目的探讨蟛蜞菊内酯是否通过调控铁死亡从而减轻高氧性急性肺损伤(HALI),为HALI的药物治疗提供理论依据。方法将24只C57BL/6J小鼠按随机数字表法分为常氧对照组、HALI模型组和蟛蜞菊内酯预处理组,每组8只。蟛蜞菊内酯预处理组经腹腔注射50 mg/kg蟛蜞菊内酯预处理6 h,其余两组不进行任何预处理;之后维持制模仓内二氧化碳含量<0.5%、氧含量>90%48 h构建HALI模型,常氧对照组置于室内空气中。制模后处死小鼠取肺组织,光镜下观察肺组织病理学改变并进行病理学评分,计算肺湿/干质量比值(W/D);检测肺组织肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-6、IL-1β)、超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)水平;采用蛋白质免疫印迹试验(Western blotting)检测肺组织谷胱甘肽过氧化物酶4(GPX4)蛋白表达。结果光镜下可见HALI模型组小鼠肺泡结构破坏,肺泡和肺间质有大量中性粒细胞浸润且肺间质增厚;肺损伤病理评分(分:0.75±0.02比0.11±0.01)和肺W/D比值(6.23±0.34比3.68±0.23)较常氧对照组明显升高(均P<0.05)。蟛蜞菊内酯预处理组小鼠肺泡腔清晰,中性粒细胞浸润和肺间质厚度不及HALI模型组,肺损伤病理评分(分:0.43±0.02比0.75±0.02)和肺W/D比值(4.56±0.12比6.23±0.34)较HALI模型组明显降低(均P<0.05)。与常氧对照组比较,HALI模型组肺组织SOD(kU/g:26.41±4.25比78.64±3.95)、GSH(mol/g:4.51±0.33比12.53±1.25)明显降低(均P<0.05),而MDA(mmol/g:54.23±4.58比9.65±1.96)、TNF-α(μg/L:96.32±3.67比11.65±2.03)、IL-6(ng/L:163.35±5.89比20.56±3.63)、IL-1β(μg/L:72.34±4.64比15.64±2.47)均明显升高(均P<0.05),GPX4蛋白表达明显降低(GPX4/β-actin:0.44±0.02比1.00±0.09,P<0.05)。与HALI模型组比较,蟛蜞菊内酯预处理组SOD(kU/g:53.28±3.69比26.41±4.25)、GSH(mol/g:9.99±0.13比4.51±0.33)明显升高(均P<0.05),MDA(mmol/g:25.36±1.98比54.23±4.58)、TNF-α(μg/L:40.25±4.13比96.32±3.67)、IL-6(ng/L:78.32±4.65比163.35±5.89)、IL-1β(μg/L:30.65±3.65比72.34±4.64)均明显降低(均P<0.05),GPX4蛋白表达明显升高(GPX4/β-actin:0.68±0.04比0.44±0.02,P<0.05)。结论蟛蜞菊内酯可通过调控铁死亡从而减轻HALI。
ObjectiveTo study whether wedelolactone can reduce hyperoxia-induced acute lung injury(HALI)by regulating ferroptosis,and provide a basic theoretical basis for the drug treatment of HALI.MethodsA total of 24 C57BL/6J mice were randomly divided into normal oxygen control group,HALI model group and wedelolactone pretreatment group,with 8 mice in each group.Mice in wedelolactone pretreatment group were treated with wedelolactone 50 mg/kg intraperitoneally for 6 hours,while the other two groups were not given with wedelolactone.After that,the HALI model was established by maintaining the content of carbon dioxide<0.5%and oxygen>90%in the molding chamber for 48 hours,and the normal oxygen control group was placed in indoor air.After modeling,the mice were sacrificed and lung tissues were collected.The lung histopathological changes were observed under light microscope and pathological scores were performed to calculate the ratio of lung wet/dry mass(W/D).The levels of tumor necrosis factor-α(TNF-α),interleukins(IL-6,IL-1β),superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione(GSH)in lung tissues of mice in each group were determined.The protein expression of glutathione peroxidase 4(GPX4)in lung tissue was detected by Western blotting.ResultsUnder light microscope,the alveolar structure of HALI model group was destroyed,and a large number of neutrophils infiltrated the alveolar and interstitial lung,and the interstitial lung was thickened.The pathological score of lung injury(score:0.75±0.02 vs.0.11±0.01)and the ratio of lung W/D(6.23±0.34 vs.3.68±0.23)were significantly higher than those in the normal oxygen control group(both P<0.05).Wedelolactone pretreated mice had clear alveolar cavity and lower neutrophil infiltration and interstitial thickness than HALI group.Pathological scores(score:0.43±0.02 vs.0.75±0.02)and W/D ratio(4.56±0.12 vs.6.23±0.34)were significantly lower than HALI group(both P<0.05).Compared with the normal oxygen control group,the levels of SOD(kU/g:26.41±4.25 vs.78.64±3.95)and GSH(mol/g:4.51±0.33 vs.12.53±1.25)in HALI group were significantly decreased,while the levels of MDA(mmol/g:54.23±4.58 vs.9.65±1.96),TNF-α(μg/L:96.32±3.67 vs.11.65±2.03),IL-6(ng/L:163.35±5.89 vs.20.56±3.63)and IL-1β(μg/L:72.34±4.64 vs.15.64±2.47)were significantly increased,and the protein expression of GPX4(GPX4/β-actin:0.44±0.02 vs.1.00±0.09)was significantly decreased(all P<0.05).Compared with the HALI group,the levels of SOD(kU/g:53.28±3.69 vs.26.41±4.25)and GSH(mol/g:6.73±0.97 vs.12.53±1.25)were significantly higher in the wedelolactone pretreatment group,and the levels of MDA(mmol/g:25.36±1.98 vs.54.23±4.58),TNF-α(μg/L:40.25±4.13 vs.96.32±3.67),IL-6(ng/L:78.32±4.65 vs.163.35±5.89),and IL-1β(μg/L:30.65±3.65 vs.72.34±4.64)were significantly lower(all P<0.05),and protein expression of GPX4 was significantly higher(GPX4/β-actin:0.68±0.04 vs.0.44±0.02,P<0.05).ConclusionWedelolactone attenuates HALI injury by regulating ferroptosis.
作者
刘君亚
覃松
冯帮海
陈淼
梅鸿
Liu Junya;Qin Song;Feng Banghai;Chen Miao;Mei Hong(Department of Critical Care Unit,Affiliated Hospital of Zunyi Medical University,Zunyi 563000,Guizhou,China;Department of Critical Care Medicine,Zunyi City Hospital of Traditional Chinese Medicine,Zunyi 563000,Guizhou,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2023年第11期1177-1181,共5页
Chinese Critical Care Medicine
基金
国家自然科学基金项目(81960362,81960024)
贵州省卫生健康委科学技术基金项目(gzwjkj2019-1-068)
贵州省科技厅自然科学项目(ZK[2022]660,ZK[2023]544)。
关键词
蟛蜞菊内酯
铁死亡
高氧性急性肺损伤
Wedelolactone
Ferroptosis
Hyperoxia-induced acute lung injury
