PDF(Pubmed)
Abstract：
OBJECTIVE: To investigate the protective effect of 5-hydroxy-6,7-dimethoxyflavone (5-HDF), a compound extracted from Elsholtzia blanda Benth., against lung injury induced by H1N1 influenza virus and explore its possible mechanism of action.
METHODS: 5-HDF was extracted from Elsholtzia blanda Benth. using ethanol reflux extraction and silica gel chromatography and characterized using NMR and MS analyses. In an A549 cell model of H1N1 influenza virus infection (MOI=0.1), the cytotoxicity of 5-HDF was assessed using MTT assay, and its effect on TRAIL and IL-8 expressions was examined using flow cytometry; Western blotting was used to detect the expression levels of inflammatory, apoptosis, and ferroptosis-related proteins. In a mouse model of H1N1 influenza virus infection established by nasal instillation of 50 μL H1N1 virus at the median lethal dose, the effects of 30 and 60 mg/kg 5-HDF by gavage on body weight, lung index, gross lung anatomy and lung histopathology were observed.
RESULTS: 5-HDF exhibited no significant cytotoxicity in A549 cells within the concentration range of 0-200 μg/mL. In H1N1-infected A549 cells, treatment with 5-HDF effectively inhibited the activation of phospho-p38 MAPK and phospho-NF-κB p65, lowered the expressions of IL-8, enhanced the expression of anti-ferroptosis proteins (SLC7A11 and GPX4), and inhibited the expressions of apoptosis markers PARP and caspase-3 and the apoptotic factor TRAIL. In H1N1-infected mice, treatment with 5-HDF for 7 days significantly suppressed body weight loss and increment of lung index and obviously alleviated lung tissue pathologies.
CONCLUSIONS: 5-HDF offers protection against H1N1 influenza virus infection in mice possibly by suppressing H1N1-induced ferroptosis, inflammatory responses, and apoptosis via upregulating SLC7A11 and GPX4, inhibiting the activation of phospho-NF-κB p65 and phospho-p38 MAPK, and decreasing the expression of cleaved caspase3 and cleaved PARP.
METHODS: 5-HDF was extracted from Elsholtzia blanda Benth. using ethanol reflux extraction and silica gel chromatography and characterized using NMR and MS analyses. In an A549 cell model of H1N1 influenza virus infection (MOI=0.1), the cytotoxicity of 5-HDF was assessed using MTT assay, and its effect on TRAIL and IL-8 expressions was examined using flow cytometry; Western blotting was used to detect the expression levels of inflammatory, apoptosis, and ferroptosis-related proteins. In a mouse model of H1N1 influenza virus infection established by nasal instillation of 50 μL H1N1 virus at the median lethal dose, the effects of 30 and 60 mg/kg 5-HDF by gavage on body weight, lung index, gross lung anatomy and lung histopathology were observed.
RESULTS: 5-HDF exhibited no significant cytotoxicity in A549 cells within the concentration range of 0-200 μg/mL. In H1N1-infected A549 cells, treatment with 5-HDF effectively inhibited the activation of phospho-p38 MAPK and phospho-NF-κB p65, lowered the expressions of IL-8, enhanced the expression of anti-ferroptosis proteins (SLC7A11 and GPX4), and inhibited the expressions of apoptosis markers PARP and caspase-3 and the apoptotic factor TRAIL. In H1N1-infected mice, treatment with 5-HDF for 7 days significantly suppressed body weight loss and increment of lung index and obviously alleviated lung tissue pathologies.
CONCLUSIONS: 5-HDF offers protection against H1N1 influenza virus infection in mice possibly by suppressing H1N1-induced ferroptosis, inflammatory responses, and apoptosis via upregulating SLC7A11 and GPX4, inhibiting the activation of phospho-NF-κB p65 and phospho-p38 MAPK, and decreasing the expression of cleaved caspase3 and cleaved PARP.
摘要：
目的：研究5-羟基-6,7-二甲氧基黄酮（5-HDF）的保护作用，从ElsholtziablandaBenth中提取的化合物。,并探讨其可能的作用机制。
方法：5-HDF从ElsholtziablandaBenth中提取。使用乙醇回流提取和硅胶色谱法，并使用NMR和MS分析进行表征。在H1N1流感病毒感染的A549细胞模型中（MOI=0.1），5-HDF的细胞毒性用MTT法评估，流式细胞术检测其对TRAIL和IL-8表达的影响;免疫印迹法检测炎症,凋亡，和铁凋亡相关蛋白。在以中位致死剂量经鼻滴注50μLH1N1病毒建立的H1N1流感病毒感染小鼠模型中，30和60mg/kg5-HDF灌胃对体重的影响，肺指数,观察肺大体解剖和肺组织病理学。
结果：5-HDF在0-200μg/mL的浓度范围内对A549细胞无明显的细胞毒性。在H1N1感染的A549细胞中，5-HDF治疗能有效抑制磷酸-p38MAPK和磷酸-NF-κBp65的活化,降低IL-8的表达,增强抗铁凋亡蛋白(SLC7A11和GPX4)的表达,并抑制凋亡标志物PARP和caspase-3以及凋亡因子TRAIL的表达。在H1N1感染的小鼠中，5-HDF治疗7天可显着抑制体重下降和肺指数升高，并明显减轻肺组织病变。
结论：5-HDF在小鼠中可能通过抑制H1N1诱导的铁细胞凋亡来提供对H1N1流感病毒感染的保护，炎症反应，通过上调SLC7A11和GPX4，抑制磷酸化-NF-κBp65和磷酸化-p38MAPK的激活，并降低裂解的caspase3和裂解的PARP的表达。
方法：5-HDF从ElsholtziablandaBenth中提取。使用乙醇回流提取和硅胶色谱法，并使用NMR和MS分析进行表征。在H1N1流感病毒感染的A549细胞模型中（MOI=0.1），5-HDF的细胞毒性用MTT法评估，流式细胞术检测其对TRAIL和IL-8表达的影响;免疫印迹法检测炎症,凋亡，和铁凋亡相关蛋白。在以中位致死剂量经鼻滴注50μLH1N1病毒建立的H1N1流感病毒感染小鼠模型中，30和60mg/kg5-HDF灌胃对体重的影响，肺指数,观察肺大体解剖和肺组织病理学。
结果：5-HDF在0-200μg/mL的浓度范围内对A549细胞无明显的细胞毒性。在H1N1感染的A549细胞中，5-HDF治疗能有效抑制磷酸-p38MAPK和磷酸-NF-κBp65的活化,降低IL-8的表达,增强抗铁凋亡蛋白(SLC7A11和GPX4)的表达,并抑制凋亡标志物PARP和caspase-3以及凋亡因子TRAIL的表达。在H1N1感染的小鼠中，5-HDF治疗7天可显着抑制体重下降和肺指数升高，并明显减轻肺组织病变。
结论：5-HDF在小鼠中可能通过抑制H1N1诱导的铁细胞凋亡来提供对H1N1流感病毒感染的保护，炎症反应，通过上调SLC7A11和GPX4，抑制磷酸化-NF-κBp65和磷酸化-p38MAPK的激活，并降低裂解的caspase3和裂解的PARP的表达。
