三白草酮通过铁死亡相关通路改善化疗药物阿霉素诱导的心肌细胞损伤
Sauchinone improves myocardial cell injury induced by chemotherapy drug doxorubicin through ferroptosis related pathway
孙星 1屈中玉 1万里新 1刘越 1赵得堡 1杜云辉 1木亚林 1刘丽娜2
作者信息
- 1. 南阳市中心医院肿瘤内科,南阳 473000
- 2. 南阳市第二人民医院肿瘤内科,南阳 473000
- 折叠
摘要
目的 探讨三白草酮(sauchinone,Sau)对化疗药物阿霉素(doxorubicin,Dox)诱导的心肌细胞损伤的改善作用及铁死亡相关机制.方法 选用心肌细胞系H9c2 细胞,首先对Sau的细胞毒性进行分析,然后检测Sau对Dox细胞毒性的影响,并用SIRT1抑制剂分析SIRT1/Nrf2信号通路在Sau影响Dox心肌细胞毒性中的作用.以CCK-8细胞活性检测,乳酸脱氢酶(LDH)、肌酸激酶同工酶MB(CK-MB)、丙二醛(MDA)及4-羟基壬烯醛(4-HNE)活性或水平的试剂盒检测,细胞内Fe2+水平的FerroOrange染色和Fe2+水平检测试剂盒检测,线粒体膜电位JC-1染色,SIRT/Nrf2信号通路蛋白和谷胱甘肽过氧化酶 4(GPX4)水平的Western blot检测等评估心肌细胞损伤程度和Sau作用机制.结果 在Sau浓度为0～10 μmol/L时,H9c2细胞活性不变;Sau以剂量依赖性改善Dox诱导的H9c2细胞活力降低;Sau对H9c2细胞线粒体膜电位水平、Fe2+浓度、脂质过氧化物水平、SIRT/Nrf2信号通路蛋白GPX4水平无明显影响;Dox诱导H9c2细胞线粒体膜电位降低、Fe2+和脂质过氧化物水平升高、SIRT1/Nrf2信号通路活性与GPX4水平显著降低,Sau对Dox诱导的上述毒性具有明显抑制作用,该抑制作用可被SIRT1/Nrf2信号通路抑制剂阻断.结论 Sau可通过激活SIRT1/Nrf2信号通路降低Dox诱导的心肌细胞铁死亡,以发挥其保护作用.
Abstract
Objective To investigate the protective effects of sauchinone(Sau)against doxorubicin(Dox)-induced cardiomyo-cyte damage and its associated mechanisms involving ferroptosis.Methods The cardiomyocyte line H9c2 was employed to assess the cytotoxicity of Sau and its impact on Dox-induced toxicity.The role of the SIRT1/Nrf2 signaling pathway was analyzed using a SIRT1 inhibitor.Cell viability was evaluated using the CCK-8 assay,while levels of lactate dehydrogenase(LDH),creatine kinase-MB isoenzyme(CK-MB),malondialdehyde(MDA),and 4-hydroxynonenal(4-HNE)were measured with respective kits.Intracellular Fe2+levels were determined using FerroOrange staining and a specific assay kit.Mitochondrial membrane potential was assessed via JC-1 staining,and Western blot was used to analyze the levels of the SIRT/Nrf2 pathway proteins and level of glutathione peroxidase 4(GPX4).Results Sau at concentrations of 0 to10 μmol/L did not affect H9c2 cell viability and dose-dependently improved the re-duction in cell viability induced by Dox.Sau did not significantly alter mitochondrial membrane potential,Fe2+concentration,lipid peroxidation levels,or the levels of the SIRT/Nrf2 pathway proteins and GPX4 in H9c2 cells.Dox-induced reductions in mitochon-drial membrane potential,increases in levels of Fe2+and lipid peroxidation,and decreases in levels of SIRT1/Nrf2 pathway proteins and GPX4 were significantly mitigated by Sau.These protective effects of Sau were blocked by the SIRT1/Nrf2 signaling pathway inhibitors.Conclusion Sau can activate the SIRT1/Nrf2 signaling pathway to reduce Dox-induced cardiomyocyte ferroptosis,thereby exerting its protective effects.
关键词
三白草酮/阿霉素/心肌毒性/SIRT1/Nrf2信号通路/铁死亡Key words
Sauchinone/doxorubicin/cardiotoxicity/SIRT1/Nrf2 signaling pathway/ferroptosis引用本文复制引用
基金项目
河南省医学科技攻关计划(联合共建)项目(LHGJ20191463)
出版年
2024
NETL
NSTL
万方数据