GDF3通过NRF2/GPX4调控铁死亡减轻脓毒症急性肺损伤
GDF3 alleviates sepsis-induced acute lung injury by regulating ferroptosis through NRF2/GPX4
王慧娟 1雷佳羲 1邹丽绢 1王璐 1刘世平 1张云龙 1詹丽英1
作者信息
- 1. 武汉大学人民医院重症医学科 湖北 武汉 430060
- 折叠
摘要
目的:探讨生长分化因子3(GDF3)通过调控铁死亡减轻小鼠脓毒症急性肺损伤的作用机制.方法:雄性C57BL/6小鼠30只,随机分成Control组、CLP组、CLP+GDF3组,每组各10只.采用盲肠结扎穿孔法(CLP)制备脓毒症肺损伤小鼠模型,CLP+GDF3组在CLP造模前尾静脉注射GDF3重组蛋白(20 μg/kg).造模24 h后取材,HE染色观察肺组织病理形态并评分,计算肺组织湿重干重比;瑞氏-吉姆萨染色分类计数各组小鼠肺泡灌洗液(BALF)中炎症细胞数量,ELISA法检测各组小鼠血清及BALF中炎性细胞因子TNF-α与IL-1β水平和肺组织中MDA、GSH与Fe2+水平,q-PCR与Western Blot法检测肺组织中GPX4、HO-1、NRF2 mRNA与蛋白质表达水平,免疫组化与免疫荧光染色检测GPX4蛋白表达.结论:与Control组相比,CLP组、CLP+GDF3组肺组织损伤明显加重,肺湿/干重比明显升高,BALF中炎性细胞数目增加,血清及BALF中TNF-α及IL-1β水平升高,肺组织中MDA与Fe2+含量增加,GSH含量降低,肺组织GPX4、HO-1与NRF2 mRNA与蛋白水平表达水平降低(P<0.05);与CLP组相比,CLP+GDF3组肺组织损伤明显减轻,肺湿/干重比下降,BALF中炎性细胞减少,血清及BALF中TNF-α及IL-1β水平下降,肺组织中MDA与Fe2+含量减少,GSH含量增加,肺组织GPX4、HO-1 与NRF2 在mRNA与蛋白水平表达升高(P<0.05).结论:GDF3 可以减轻脓毒症小鼠急性肺损伤,作用机制可能与激活NRF2/GPX4信号通路、抑制铁死亡相关.
Abstract
Objective:To investigate the mechanism of growth differentiation factor-3(GDF3)in alleviating sepsis-induced acute lung injury in mice by regulating ferroptosis.Methods:Thirty male C57BL/6 mice were randomly divided into three groups:the control group,the cecal ligation and puncture(CLP)CLP group,and the CLP+GDF3 group,with 10 mice in each group.The septic lung injury mouse model was established using the CLP method.In the CLP+GDF3 group,GDF3(20 μg/kg)was administered via tail vein injection before CLP modeling.Tissue samples were collected 24 hours af-ter modeling.Lung tissue pathological morphology was observed and scored through HE staining,and the wet-to-dry weight ratio of lung tissue was calculated.The inflammatory cell count in the bron-choalveolar lavage fluid(BALF)of each group of mice was assessed using Wright's Giemsa staining method.The levels of inflammatory cytokines TNF-α and IL-1β in the serum and BALF of each group,as well as the levels of MDA,GSH,and Fe2+in lung tissue,were measured by ELISA.GPX4,HO-1,and NRF2 expression levels at the mRNA and protein levels in lung tissue were evalu-ated using q-PCR and Western Blot.GPX4 expression was detected using immunohistochemistry and immunofluorescence staining.Results:Compared with the control group,the CLP group exhibited significant exacerbation of lung tissue injury,a notable increase in the wet-to-dry weight ratio of the lungs,an elevated count of inflammatory cells in BALF,increased levels of TNF-α and IL-1β in both serum and BALF,elevated levels of MDA and Fe2+in lung tissue,decreased GSH content,and a downregulation of GPX4,HO-1 and NRF2 expression at both mRNA and protein levels in lung tis-sue(P<0.05).In comparison to the CLP group,the CLP+GDF3 group demonstrated significant at-tenuation of lung tissue injury,including a remarkable decrease in the wet-to-dry weight ratio,a reduc-tion in the count of inflammatory cells in BALF,decreased levels of TNF-α and IL-1β in both serum and BALF,decreased levels of MDA and Fe2+,increased GSH content,and upregulation of GPX4,HO-1 and NRF2 expression at both mRNA and protein levels in lung tissue(P<0.05).Conclusion:GDF3 can improve CLP-induced septic ALI in mice,and its mechanism of action may be related to the activation of the NRF2/GPX4 signaling pathway and the inhibition of ferroptosis.
关键词
脓毒症/急性肺损伤/GDF3/铁死亡/NRF2/GPX4Key words
Sepsis/Acute Lung Injury/GDF3/Ferroptosis/NRF2/GPX4引用本文复制引用
基金项目
国家重点研发项目(2021YFC2501800)
国家自然科学基金青年基金资助项目(82102245)
出版年
2024
NETL
NSTL
万方数据