黄芩苷通过调控铁死亡减轻H9c2细胞缺氧复氧损伤及其机制研究  

Effect and Mechanism of Baicalin on Hypoxia-Reoxygenation Injury in H9c2 Cells Via Regulating Ferroptosis

作　　者：李家权 许锦 谭子富 方孝俊 LI Jiaquan;XU jin;TAN Zifu;FANG Xiaojun(The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture,Enshi Hubei 445000,China;The Central Hospital of Enshi,Enshi Hubei 445000,China)

机构地区：[1]恩施土家族苗族自治州中心医院,湖北恩施445000 [2]恩施市中心医院,湖北恩施445000

出　　处：《中医药导报》2025年第2期13-18,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖北省中医药管理局科研项目(ZY2023F121)。

摘　　要：目的:研究黄芩苷通过铁死亡途径对H9c2心肌细胞缺氧复氧损伤的作用及机制。方法:采用Na_(2)S_(2)O_(4)诱导大鼠H9c2心肌细胞铁死亡模型,随后用黄芩苷干预。采用试剂盒检测细胞活力、ROS、线粒体膜电位和Fe^(2+)水平。Western blotting检测细胞PTGS2、NOX2和GPX4蛋白表达水平。PTGS2激动剂BUR1联合黄芩苷干预Na_(2)S_(2)O_(4)诱导的细胞后,试剂盒检测细胞Fe^(2+)水平。结果:80μmol/L以内的黄芩苷对H9c2细胞无毒性。Na_(2)S_(2)O_(4)诱导H9c2细胞铁死亡,表现为细胞ROS和Fe^(2+)水平升高,线粒体膜电位坍塌,GPX4表达明显下降(P<0.05)。黄芩苷干预后,Na_(2)S_(2)O_(4)诱导的细胞ROS和Fe^(2+)水平下降,线粒体膜电位恢复,GPX4表达明显升高(P<0.05),铁死亡进程被逆转。Na_(2)S_(2)O_(4)诱导H9c2细胞后,PTGS2和NOX4表达明显升高(P<0.05),而黄芩苷干预后,Na_(2)S_(2)O_(4)诱导的H9c2心肌细胞PTGS2和NOX4表达明显降低(P<0.05)。黄芩苷与PTGS2和NOX4有很好的结合活性,结合能分别为-10.65和-8.82 kcal/mol。BUR1能够逆转黄芩苷抑制的Fe^(2+)水平。结论:黄芩苷对Na_(2)S_(2)O_(4)诱导H9c2细胞建立的缺氧复氧损伤模型铁死亡具有明确的恢复作用,其机制可能与其抑制PTGS2和NOX4蛋白表达有关。Objective:To investigate the effect and mechanism of baicalin on hypoxia-reoxygenation injury in H9c2 cells via regulating ferroptosis.Methods:Rat H9c2 cardiomyocytes hypoxia-reoxygenation injury model was constructed with Na_(2)S_(2)O_(4),followed by baicalin intervention.Cell viability,ROS,mitochondrial membrane potential,and Fe^(2+)levels were measured using assay kits.Western blotting was utilized to evaluate cell PTGS2,NOX2,and GPX4 protein expression levels.Cell Fe^(2+)levels were examined using assay kits after baicalin intervention with PTGS2 agonist BUR1 following Na_(2)S_(2)O_(4)induction.Results:Baicalin up to 80μmol/L showed no toxicity to H9c2 cells.Na_(2)S_(2)O_(4)induced ferroptosis in H9c2 cells,characterized by elevated ROS and Fe^(2+)levels,mitochondrial membrane potential collapse,and significant decrease in GPX4 expression(P<0.05).Baicalin intervention led to decreased ROS and Fe^(2+)levels induced by Na_(2)S_(2)O_(4),restored mitochondrial membrane potential,markedly increased GPX4 expression(P<0.05),and reversing the process of ferroptosis.After Na_(2)S_(2)O_(4)induction in H9c2 cells,PTGS2 and NOX4 expression significantly increased(P<0.05),while baicalin markedly reduced the expression of PTGS2 and NOX4 induced by Na_(2)S_(2)O_(4)(P<0.05).Baicalin exhibited good binding activity with PTGS2 and NOX4,with binding energies of-10.65 and-8.82 kcal/mol,respectively.BUR1 could reverse the baicalin-inhibited Fe^(2+)levels.Conclusion:Baicalin demonstrates a clear therapeutic effect on ferroptosis in the hypoxia-reoxygenation injury model established by Na_(2)S_(2)O_(4)-induced H9c2 cells,and its treatment mechanism may be associated with the inhibition of PTGS2 and NOX4 protein expression.

关 键 词：缺血性心肌病 黄芩苷 铁死亡 大鼠H9c2心肌细胞 Na_(2)S_(2)O_(4) PTGS2 NOX4 

分 类 号：R285.5[医药卫生—中药学]