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

作　　者：孙星 屈中玉[1] 万里新[1] 刘越[1] 赵得堡 杜云辉 木亚林[1] 刘丽娜[2] Sun Xing;Qu Zhongyu;Wan Lixin;Liu Yue;Zhao Debao;Du Yunhui;Mu Yalin;Liu Lina(Department of Medical Oncologist,Nanyang Central Hospital,Nanyang 473000;Department of Medical Oncologist,Nanyang Second General Hospital,Nanyang 473000)

机构地区：[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

出　　处：《中国组织化学与细胞化学杂志》2024年第2期118-125,共8页Chinese Journal of Histochemistry and Cytochemistry

基　　金：河南省医学科技攻关计划(联合共建)项目(LHGJ20191463)。

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Fe^(2+)水平的FerroOrange染色和Fe^(2+)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0~10μmol/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、Fe^(2+)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、Fe^(2+)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。Objective To investigate the protective effects of sauchinone(Sau)against doxorubicin(Dox)-induced cardiomyocyte 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 Fe^(2+)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 to10μmol/L did not affect H9c2 cell viability and dose-dependently improved the reduction in cell viability induced by Dox.Sau did not significantly alter mitochondrial membrane potential,Fe^(2+)concentration,lipid peroxidation levels,or the levels of the SIRT/Nrf2 pathway proteins and GPX4 in H9c2 cells.Dox-induced reductions in mitochondrial membrane potential,increases in levels of Fe^(2+)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 

分 类 号：R730.5[医药卫生—肿瘤]