Erastin通过MAPK介导的氧化应激信号通路诱导肺成纤维细胞铁死亡
Erastin induces ferroptosis in lung fibroblasts through MAPK mediated oxidative stress signaling pathway
王沂然 1张诗捷 1关予博 2李渺渺 2蔡汝忆 3武琦1
作者信息
- 1. 徐州医科大学生理学教研室,徐州 221004
- 2. 徐州医科大学病理生理学教研室,徐州 221004
- 3. 徐州医科大学第一临床医学院,徐州 221004
- 折叠
摘要
目的 探究Erastin对肺成纤维细胞铁死亡的作用机制.方法 向小鼠肺成纤维细胞(C57/B6-L)中加入不同浓度的铁死亡诱导剂Erastin,通过细胞计数试剂(CCK-8)检测细胞活性、荧光显微镜观察氧化应激水平,蛋白质印迹法(Western blot)检测丝裂原活化蛋白激酶(MAPK)信号通路相关蛋白表达,之后分别加入p38和人细胞外信号调节激酶(ERK)的抑制剂SB203580和U0126,进一步验证Erastin诱导肺成纤维细胞铁死亡的作用机制.结果 Erastin在100μmol/L药物浓度时,能够明显诱导肺成纤维细胞铁死亡,同时伴有氧化应激表达增强,此外MAPK通路中的p38和ERK蛋白磷酸化水平升高(P<0.05).而在加入SB203580、U0126抑制剂后,肺成纤维细胞氧化应激水平明显下降,同时细胞的活性明显升高(P<0.05).结论 Erastin能够诱导肺成纤维细胞铁死亡,其作用机制可能与MAPK信号通路介导的氧化应激有关.
Abstract
Objective To investigate the mechanism by which Erastin affects ferroptosis in lung fibroblasts.Meth-ods Mouse lung fibroblasts (C57/B6-L) were treated with varying concentrations of the iron death inducer Eras-tin.Cell viability was assessed using the cell counting Kit-8 (CCK-8) assay.Oxidative stress levels were visualized using a fluorescence microscope, and the expression of proteins related to the mitogen-activated protein kinase (MAPK) signaling pathway was analyzed using Western blot.Additionally, the p38 and extracellular regulated protein kinase (ERK) inhibitors SB203580 and U0126 were employed to further elucidate the mechanism by which Erastin induces iron death in lung fibroblasts.Results At a concentration of 100 μmol/L, Erastin effectively in-duced ferroptosis in lung fibroblasts, leading to an upregulation of oxidative stress.Furthermore, the phosphoryla-tion levels of p38 and ERK proteins in the MAPK pathway were elevated (P<0.05) .The addition of SB203580 and U0126 inhibitors resulted in a significant reduction in oxidative stress levels and a notable increased in cell ac-tivity in lung fibroblasts (P<0.05).Conclusion It can be concluded that Erastin induces ferroptosis in lung fi-broblasts, potentially through the mediation of oxidative stress via the MAPK signaling pathway.
关键词
肺纤维化/肺成纤维细胞/铁死亡/活性氧/MAPK信号通路Key words
pulmonary fibrosis/lung fibroblasts/ferroptosis/reactive oxygen species/MAPK signaling pathway引用本文复制引用
基金项目
国家自然科学基金(82104624)
江苏省高等学校自然科学研究面上项目(21KJB320003)
江苏省高校大学生创新创业重点项目(202210313048Z)
出版年
2024
国家科技期刊平台
NSTL
万方数据