芦荟素减轻大鼠心肌细胞缺氧损伤:抑制氧化应激和铁死亡  

Aloin mitigates hypoxic injury in rat cardiomyocytes:inhibiting oxidative stress and ferroptosis

作　　者：谭铭月 金奕丰 张俊[1] 李红霞[1] Tan Mingyue;Jin Yifeng;Zhang Jun;Li Hongxia(Department of Cardiology,First Affiliated Hospital of Soochow University,Suzhou 215006,Jiangsu Province,China;Department of General Medicine,First Affiliated Hospital of Soochow University,Suzhou 215006,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院心血管内科,江苏省苏州市215006 [2]苏州大学附属第一医院全科医学科,江苏省苏州市215006

出　　处：《中国组织工程研究》2025年第25期5335-5344,共10页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81100173),项目负责人:李红霞。

摘　　要：背景:心肌细胞缺氧损伤与氧化应激和铁死亡密切有关。既往研究表明芦荟素具有抗氧化、抗炎、抗肿瘤等多种作用。目的:探讨芦荟素对缺氧诱导的H9C2细胞氧化应激及铁死亡的影响。方法:利用H9C2细胞构建缺氧模型,首先通过检测细胞活性,确定芦荟素无细胞毒性及最佳干预浓度。随后,利用试剂盒、超氧化物阴离子荧光探针及MitoSOX^(TM)Red荧光探针,分别评估芦荟素对缺氧诱导的H9C2细胞乳酸脱氢酶释放、活性氧产生以及线粒体氧化应激的影响。为了验证芦荟素对铁死亡的影响,采用荧光染色和流式细胞术,分别检测细胞内Fe^(2+)的含量和脂质过氧化程度。接着,利用蛋白免疫印迹和实时荧光定量PCR技术,检测铁死亡调节因子(谷胱甘肽过氧化物酶4、酰基辅酶A合成酶长链家族成员4和核因子E2相关因子2)的表达。最后,通过运用铁死亡诱导剂Erastin,进一步确认铁死亡在芦荟素介导心肌保护过程中的重要作用。结果与结论:(1)与对照组相比,缺氧组H9C2细胞活力显著降低,乳酸脱氢酶释放、活性氧水平、线粒体氧化应激程度、Fe^(2+)含量及脂质过氧化程度显著上升,谷胱甘肽过氧化物酶4、核因子E2相关因子2的mRNA和蛋白表达明显降低,酰基辅酶A合成酶长链家族成员4的mRNA和蛋白表达显著升高(均P<0.05);(2)与缺氧组比较,芦荟素低、高剂量组逆转了上述指标变化(均P<0.05);(3)与缺氧+芦荟素组相比,缺氧+芦荟素+Erastin组H9C2细胞活力明显下降,乳酸脱氢酶释放显著升高(均P<0.01)。结果表明,芦荟素对缺氧处理的H9C2细胞具有保护作用,且呈剂量依赖性,主要通过抑制氧化应激和铁死亡实现的。BACKGROUND:Myocardial cell hypoxic injury is closely associated with oxidative stress and ferroptosis.Previous studies have shown that aloin has various effects such as antioxidant,anti-inflammatory,and anti-tumor activities.OBJECTIVE:To investigate the effects of aloin on oxidative stress and ferroptosis in hypoxia-induced H9C2 cells.METHODS:A hypoxia model was established using H9C2 myocardial cells.Firstly,cell viability was determined to confirm the lack of cytotoxicity of aloin and to determine its optimal therapeutic concentration.Subsequently,the effects of aloin on hypoxia-induced lactate dehydrogenase release,reactive oxygen species production,and mitochondrial oxidative stress in H9C2 cells were evaluated using assay kits,dihydroethidium fluorescent probes,and MitoSOX^(TM)Red fluorescent probes,respectively.To verify the effect of aloin on ferroptosis,intracellular Fe^(2+)content and lipid peroxidation level were detected using fluorescence staining and flow cytometry,respectively.Then,the expression levels of ferroptosis regulatory factors glutathione peroxidase 4,acyl-CoA synthetase long-chain family member 4,and nuclear factor E2-related factor 2 were detected using western blot assay and real-time fluorescence quantitative PCR techniques.Finally,the role of ferroptosis in aloin-mediated myocardial protection was further confirmed by using the ferroptosis inducer Erastin.RESULTS AND CONCLUSION:(1)Compared with the control group,the viability of H9C2 cells in the hypoxia group was significantly decreased,lactate dehydrogenase release,reactive oxygen species level,mitochondrial oxidative stress degree,Fe^(2+)content,and lipid peroxidation degree were significantly increased,while glutathione peroxidase 4 and nuclear factor E2-related factor 2 mRNA and protein expression levels were significantly decreased,and acyl-CoA synthetase long-chain family member 4 mRNA and protein expression were significantly increased(all P<0.05).(2)Compared with the hypoxia group,both low and high doses of aloin reversed the

关 键 词：芦荟素 心肌细胞缺氧损伤 氧化应激 铁死亡 线粒体 

分 类 号：R459.9[医药卫生—治疗学] R336[医药卫生—临床医学] R684