CYP1B1抑制剂在心力衰竭细胞模型中缓解铁死亡的机制研究  

Mechanism of CYP1B1 inhibitor alleviating ferroptosis in heart failure cellular model

作　　者：王力 苏忆玲 周嘉彬 俞涛 陈楚[1] 陆齐[1] 史嘉玉 WANG Li;SU Yiling;ZHOU Jiabin;YU Tao;CHEN Chu;LU Qi;SHI Jiayu(Department of Cardiology,the Affiliated Hospital of Nantong University,Nantong 226001)

机构地区：[1]南通大学附属医院心血管内科,南通226001

出　　处：《南通大学学报（医学版）》2023年第2期122-127,共6页Journal of Nantong University(Medical sciences)

基　　金：江苏省研究生科研与实践创新计划项目(SJCX20_1154);南通市科技项目(JC2021140)。

摘　　要：目的:探讨细胞色素P4501B1(cytochrome P4501B1,CYP1B1)及其抑制剂对小鼠心肌细胞铁死亡的影响及其潜在的分子机制。方法:首先,对正常和心力衰竭小鼠行RNA测序检测差异表达的基因,使用实时荧光定量PCR(quantitative real-time PCR,qPCR)检测比较去氧肾上腺素(phenylephrine,PE)刺激组和正常组原代心肌细胞中CYP1B1的表达情况,通过丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)、Fe2+测定分析两组铁死亡指标;然后,使用不同浓度的CYP1B1抑制剂α-萘黄酮预处理心肌细胞后再行PE刺激,与对照组、PE组比较铁死亡指标,并行qPCR检测比较各组心力衰竭标志物的基因表达水平;最后,通过Western Blot和免疫荧光测定铁死亡相关蛋白核因子E2相关因子2(nuclear factor-erythroid 2-related factor 2,Nrf2)的表达。结果:PE刺激增加了原代心肌细胞的CYP1B1表达水平和铁死亡指标,而CYP1B1抑制剂α-萘黄酮能逆转这种改变,并回调了Nrf2的表达,缓解了心力衰竭基因标志物的上调。结论:CYP1B1抑制剂在心力衰竭的细胞模型中缓解了心肌细胞铁死亡,这种保护作用可能通过Nrf2实现,提示CYP1B1可能作为心力衰竭的潜在治疗靶标之一。Objective:To investigate the effect of cytochrome P4501B1(CYP1B1)and its inhibitor on ferroptosis in mouse cardiomyocytes and its underlying molecular mechanism.Methods:Firstly,RNA sequencing was performed on normal and heart failure mice to detect differentially expressed genes,quantitative real-time PCR(qPCR)was used to compare the expression of CYP1B1 in primary cardiomyocytes of the phenylephrine(PE)-stimulated group and normal group.Malondialdehyde(MDA),glutathione(GSH)and Fe2+were determined to analyze the indicators of ferroptosis in those two groups;after that,different concentrations of CYP1B1 inhibitorα-naphthoflavone was used to pretreat cardiomyocytes and then stimulated with PE.The ferroptosis indexes were compared between the control group and PE group,and the gene expression levels of heart failure markers in each group were compared by parallel qPCR detection.Finally,the expression of nuclear factor-erythroid 2-related factor 2(Nrf2)was determined by Western Blot and immunofluorescence.Results:PE stimulation increased CYP1B1 expression levels and ferroptosis markers in primary cardiomyocytes,while the CYP1B1 inhibitorα-naphthoflavone reversed these changes,rise Nrf2 expression,and alleviated the up-regulation of heart failure gene markers.Conclusion:CYP1B1 inhibitors alleviate cardiomyocyte ferroptosis in a cellular model of heart failure,and this protective effect may be achieved through Nrf2,suggesting that CYP1B1 may be one of the potential therapeutic targets for heart failure.

关 键 词：心力衰竭 细胞色素P4501B1 铁死亡 核因子E2相关因子2 α-萘黄酮 RNA测序 小鼠 

分 类 号：R541.6[医药卫生—心血管疾病]