基于Nrf2/HO-1/GPX4信号通路探讨三七皂苷R1对低压缺氧诱导的高原肺水肿的保护作用  

Investigating the Protective Effect of Notoginsenoside R1 on Hypobaric Hypoxia-Induced High-Altitude Pulmonary Edema Based on Nrf2/HO-1/GPX4 Signaling Pathway

作　　者：裴彩霞 刘俊伶 贾楠 何亚聪 王振兴 王飞[1] PEI Caixia;LIU Junling;JIA Nan;HE Yacong;WANG Zhenxing;WANG Fei(Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China;Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学附属医院,成都610075 [2]成都中医药大学,成都611137

出　　处：《世界科学技术-中医药现代化》2024年第12期3209-3217,共9页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会青年科学基金项目(82205043):麝香经RIPK3/HIF-1α信号轴“开玄通窍”调控高原脑水肿中枢细胞炎症的机制研究:聚焦小胶质细胞极化驱动坏死性凋亡的表型串扰,负责人:何亚聪;四川省中医药管理局科学技术研究专项课题(2023MS555):益气升陷法经ACE2/Ang(1-7)/MasR-胞葬轴调控高原肺水肿的机制研究,负责人:王振兴。

摘　　要：目的基于核因子红系2相关因子2(Nrf2)/血红素氧合酶1(HO-1)/谷胱甘肽过氧化物酶4(GPX4)信号通路探讨三七皂苷R1对低压缺氧诱导的高原肺水肿的干预作用及分子机制。方法将42只6周龄雄性SD大鼠适应性喂养1周后,随机分为空白组(Control)、阴性对照组(NGR1,100 mg·kg^(-1))、模型组(HH)、三七皂苷R1低剂量组(NGR1-low,50 mg·kg^(-1))、三七皂苷R1高剂量组(NGR1-high,100 mg·kg^(-1))及地塞米松组(Dex,4 mg·kg^(-1)),每组7只。给药组用三七皂苷R1或地塞米松腹腔注射给药后,使用低压缺氧模拟舱模拟高原环境进行造模。检测支气管肺泡灌洗液(BALF)中的总蛋白浓度;测量肺湿/干重比(W/D);ELISA法检测BALF中肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)和IL-1β水平;苏木素-伊红(HE)染色观察各组肺病理形态变化;生化试剂盒检测肺组织中活性氧(ROS)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)含量及活性;蛋白免疫印迹法(Western blot)检测肺组织中Nrf2、HO-1、GPX4、SLC7A11、NOX1蛋白表达水平。结果与Control组相比,HH组大鼠BALF中总蛋白浓度显著升高(P<0.01),肺W/D显著升高(P<0.01);肺组织病理显示肺泡壁明显增厚,肺间质水肿,肺泡腔、肺泡间隔及肺间质内可见出血及大量炎性渗出;BALF中TNF-α(P<0.001)、IL-6(P<0.01)和IL-1β(P<0.0001)水平显著升高;肺组织中ROS含量显著升高(P<0.01),SOD(P<0.01)、GSH(P<0.01)活性显著降低;肺组织细胞核中Nrf2的表达显著降低(P<0.001),细胞质中Nrf2的表达显著升高(P<0.05),HO-1(P<0.01)、GPX4(P<0.01)、SLC7A11(P<0.05)的表达显著降低,NOX1的表达显著升高(P<0.01)。与HH组相比,各药物干预组大鼠BALF中总蛋白浓度显著降低,肺W/D显著降低;肺组织病理明显改善;BALF中TNF-α、IL-6和IL-1β水平显著降低;肺组织中ROS含量显著降低,SOD、GSH活性显著升高;肺组织细胞核中Nrf2的表达显著升高,细胞质中Nrf2的表达显著降低,HO-1、GPX4、SLC7A1Objective To investigate the effects and molecular mechanism of Notoginsenoside R1 on hypobaric hypoxia-induced high-altitude pulmonary edema based on nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)/glutathione peroxidase 4(GPX4)signaling pathway.Methods Forty-two 6-week-old male SD rats were acclimatized and fed for one week,and then randomized into the Control group,negative control group(NGR1,100 mg·kg^(-1)),model group(HH),Notoginsenoside R1 low dose group(NGR1-low,50 mg·kg^(-1)),Notoginsenoside R1 high dose group(NGR1-high,100 mg·kg^(-1)),and dexamethasone group(Dex,4 mg·kg^(-1)),with seven rats in each group.After the drug administration group was administered with Notoginsenoside R1 or dexamethasone by intraperitoneal injection,the hypobaric hypoxia simulation chamber was used to simulate the plateau environment for modeling.Detect the total protein concentration in bronchoalveolar lavage fluid(BALF)and measure the lung wet/dry weight ratio(W/D);ELISA to measure inflammation levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1βin BALF;hematoxylin-eosin(HE)staining to visualize pathomorphologic changes in the lungs of each group;biochemical kit detect the content and activity of reactive oxygen species(ROS),superoxide dismutase(SOD),and reduced glutathione(GSH)in the lung tissues;Western blot to detect the protein expression of Nrf2,HO-1,GPX4,SLC7A11 and NOX1 in lung tissues.Results Compared with the Control group,the total protein concentration in the BALF of rats in the HH group was markedly increased(P<0.01),and the lung W/D was markedly increased(P<0.01);lung histopathology showed obvious thickening of the alveolar wall,interstitial edema,hemorrhage and massive inflammatory exudation in the alveolar lumen,alveolar septa,and interstitium of the lungs;the levels of TNF-α(P<0.001),IL-6(P<0.01),and IL-1β(P<0.0001)in the BALF were significantly elevated;ROS content was significantly increased(P<0.01),and SOD(P<0.01)and GSH(P<0.01)activities were significantl

关 键 词：高原肺水肿 NRF2 三七皂苷R1 铁死亡 氧化应激 

分 类 号：R285.5[医药卫生—中药学]