基于Nrf2/GPX4铁死亡途径探讨加味桃仁承气汤对机械通气相关性肺损伤大鼠的保护作用  

Exploring the Protective Effect of Modified Taoren Chengqi Decoction on Ventilator-induced Lung Injury in Rats Based on the Nrf2/GPX4-ferroptosis Pathway

作　　者：孙治霞 王丽辉 索红亮 李华 陈乾 SUN Zhixia;WANG Lihui;SUO Hongliang;LI Hua;CHEN Qian(Department of Critical Care Medicine,Henan Provincial Hospital of Traditional Chinese Medicine(the Second Affiliated Hospital of Henan University of Chinese Medicine),Zhengzhou 450002 Henan,China;Department of Cardiology,Henan Provincial Hospital of Traditional Chinese Medicine(the Second Affiliated Hospital of Henan University of Chinese Medicine),Zhengzhou 450002 Henan,China;Department of Emergency,Henan Provincial Hospital of Traditional Chinese Medicine(the Second Affiliated Hospital of Henan University of Chinese Medicine),Zhengzhou 450002 Henan,China)

机构地区：[1]河南省中医院(河南中医药大学第二附属医院)重症医学科,河南郑州450002 [2]河南省中医院(河南中医药大学第二附属医院)心病科,河南郑州450002 [3]河南省中医院(河南中医药大学第二附属医院)急诊科,河南郑州450002

出　　处：《中药新药与临床药理》2024年第9期1281-1288,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：河南省卫生健康委国家中医临床研究基地科研专项(2021JDZX2033)。

摘　　要：目的基于核因子E_(2)相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)铁死亡途径探讨加味桃仁承气汤对机械通气相关性肺损伤(ventilator-induced lung injury,VILI)大鼠的保护作用。方法将大鼠随机分为对照组、模型组、加味桃仁承气汤组、ML385(Nrf2抑制剂)组、加味桃仁承气汤+ML385组,每组12只。对照组大鼠进行气管插管,但自主呼吸;其他组大鼠均需进行机械通气4 h。机械通气前7 d进行给药处理,每日1次,连续7 d。机械通气结束后,ELISA法检测肺泡灌洗液(BALF)中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平;检测大鼠肺组织湿质量/干质量比值变化;检测肺组织病理;试剂盒检测大鼠肺组织中谷胱甘肽(GSH)、丙二醛(MDA)、Fe2+含量;免疫荧光染色法检测肺组织中活性氧(ROS)、4-羟基壬烯醛(4-HNE)相对荧光强度;检测肺组织中质载体家族7成员11(SLC7A11)、Nrf2、GPX4 mRNA及蛋白表达。结果与对照组比较,模型组大鼠肺组织破坏明显,BALF中TNF-α、IL-6水平升高,肺组织湿质量/干质量比值、MDA、Fe2+含量及ROS、4-HNE相对荧光强度升高,GSH含量、Nrf2、SLC7A11、GPX4 mRNA及蛋白表达降低(P<0.01)。与模型组比较,加味桃仁承气汤组大鼠肺组织病理损伤有所改善,BALF中TNF-α、IL-6水平降低,肺组织湿质量/干质量比值、MDA、Fe^(2+)含量及ROS、4-HNE相对荧光强度降低,GSH含量、Nrf2、SLC7A11、GPX4 mRNA及蛋白表达升高(P<0.01);ML385组对应指标变化趋势与上述相反(P<0.01);加味桃仁承气汤+ML385组肺组织病理损伤减轻,BALF中TNF-α、IL-6水平降低,肺组织湿质量/干质量比值、MDA、Fe2+含量及ROS、4-HNE相对荧光强度降低,GSH含量、Nrf2、SLC7A11、GPX4 mRNA及Nrf2、GPX4蛋白表达升高(P<0.01,P<0.05)。与ML385组比较,加味桃仁承气汤+ML385组肺组织病理损伤有所缓解,BALF中TNF-α、IL-6水平降低,肺组织湿质量/干质量比值、MDA、Fe2+含量及ROS、4-HNE相对荧光强度�Objective To explore the protective effect of modified Taoren Chengqi Decoction on ventilator-induced lung injury(VILI)in rats based on the nuclear factor-erythroid 2-related factor 2(Nrf2)/glutathione peroxidase 4(GPX4)-ferroptosis pathway.Methods Rats were randomly separated into the control group,the model group,the modified Taoren Chengqi Decoction group,the ML385(Nrf2 inhibitor)group,and the modified Taoren Chengqi Decoction+ML385 group,with 12 rats in each group.The rats in control group underwent tracheal intubation and kept spontaneous breathing.The rats of other groups were subjected to mechanical ventilation for 4 hours.Seven days before mechanical ventilation,medication treatment was carried out once a day for seven days.After mechanical ventilation,ELISA was applied to detect the levels of tumor necrosis factor-α(TNF-α)and interleukin 6(IL-6)in bronchoalveolar lavage fluid(BALF).Lung wet/dry weight ratio and lung tissue pathology of rat were detected.The reagent kit was applied to detect the content of glutathione(GSH),malonaldehyde(MDA),and Fe^(2+)in rat lung tissue.The relative fluorescence intensity of reactive oxygen species(ROS)and 4-hydroxynonenal(4-HNE)in lung tissue was detected by immunofluorescence staining.The mRNA and protein expressions of solute carrier family 7 member 11(SLC7A11),Nrf2,GPX4 were detected.Results Compared with the control group,the lung tissue of rats in model group was severely damaged,the levels of TNF-αand IL-6 in BALF increased,lung wet/dry weight ratio,content of MDA and Fe^(2+),and the relative fluorescence intensity of ROS and 4-HNE increased,but the content of GSH,the mRNA and protein expressions of Nrf2,SLC7A11,and GPX4 decreased(P<0.01).Compared with the model group,the pathological damage of lung tissue in the modified Taoren Chengqi Decoction group was improved,the levels of TNF-αand IL-6 in BALF decreased,lung wet/dry weight ratio,content of MDA and Fe^(2+),the relative fluorescence intensity of ROS and 4-HNE decreased,the content of GSH,the mRNA and prot

关 键 词：加味桃仁承气汤 机械通气相关性肺损伤 核因子E2相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)通路 铁死亡 炎症 大鼠 

分 类 号：R285.5[医药卫生—中药学]