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双硫仑通过抑制TAK1介导的PANoptosis减轻心肌肥大损伤

李卫东^*, 沈玄洋, 蒋晓璐, 文红福, 沈媛, 张美琪, 谭文涛

川北医学院附属医院急诊科，南充 637000

摘要

本文旨在探讨双硫仑(disulfiram, DSF)对心肌肥大损伤的影响和潜在机制，重点研究转化生长因子β激活激酶1 (transforming growth factor-β-activated kinase 1, TAK1)介导的泛凋亡(PANoptosis)的作用。培养H9C2心肌细胞，用血管紧张素II (angiotensin II, Ang II, 1 μmol/L)处理，建立心肌肥大损伤模型。DSF (40 μmol/L)单独或联合TAK1抑制剂5z-7-oxozeaenol(5z-7, 0.1 μmol/L)处理该心肌细胞肥大损伤模型。用碘化丙啶(propidium iodide, PI)染色法评估细胞损伤，用CCK8法检测细胞活力，用ELISA检测细胞培养液中炎症因子水平，用Western blot检测TAK1和PANoptosis通路关键蛋白的表达水平，用免疫共沉淀检测TAK1和RIPK1结合率，用鬼笔环肽染色法检测心肌细胞表面积。结果显示，Ang II显著降低H9C2心肌细胞活力以及TAK1和RIPK1的结合率，显著提高H9C心肌细胞表面积、PI染色阳性率、细胞培养液中炎症因子[白细胞介素1β (interleukin-1β, IL-1β)、IL-18 和肿瘤坏死因子α (tumor necrosis factor α, TNF-α)]水平和p-TAK1/TAK1 比值，并显著上调PANoptosis通路关键蛋白[焦亡相关蛋白NLRP3、Caspase-1 (p20)和GSDMD-N (p30)，凋亡相关蛋白Caspase-3 (p17)、Caspase-7 (p20)和Caspase-8 (p18)，以及坏死性凋亡相关蛋白p-MLKL、RIPK1和RIPK3]表达水平。DSF可显著逆转Ang II诱导的以上变化。5z-7和外源性IL-1β均可削弱DSF的这些心肌保护作用。以上结果提示，DSF可能通过抑制TAK1 介导的PANoptosis 减轻心肌肥大损伤。

关键词： 双硫仑; 心肌肥大; TAK1; PANoptosis; H9C2心肌细胞

Disulfiram alleviates cardiac hypertrophic injury by inhibiting TAK1-mediated PANoptosis

LI Wei-Dong^*, SHEN Xuan-Yang, JIANG Xiao-Lu, WEN Hong-Fu, SHEN Yuan, ZHANG Mei-Qi, TAN Wen-Tao

Department of Emergency, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China

Abstract

The study aims to examine the effects and potential mechanisms of disulfiram (DSF) on cardiac hypertrophic injury, focusing on the role of transforming growth factor-β-activated kinase 1 (TAK1)-mediated pan-apoptosis (PANoptosis). H9C2 cardiomyocytes were treated with angiotensin II (Ang II, 1 μmol/L) to establish an in vitro model of myocardial hypertrophy. DSF (40 μmol/L) was used to treat cardiomyocyte hypertrophic injury models, either along or in combination with the TAK1 inhibitor, 5z-7-oxozeaenol (5z-7, 0.1 μmol/L). We assessed cell damage using propidium iodide (PI) staining, measured cell viability with CCK8 assay, quantified inflammatory factor levels in cell culture media via ELISA, detected TAK1 and RIPK1 binding rates using immunoprecipitation, and analyzed the protein expression levels of key proteins in the TAK1-mediated PANoptosis pathway using Western blot. In addition, the surface area of cardiomyocytes was measured with Phalloidin staining. The results showed that Ang II significantly reduced the cellular viability of H9C2 cardiomyocytes and the binding rate of TAK1 and RIPK1, significantly increased the surface area of H9C2 cardiomyocytes, PI staining positive rate, levels of inflammatory factors [interleukin-1β (IL-1β), IL-18, and tumor necrosis factor α (TNF-α)] in cell culture media and p-TAK1/TAK1 ratio, and significantly up-regulated key proteins in the PANoptosis pathway [pyroptosis-related proteins NLRP3, Caspase-1 (p20), and GSDMD-N (p30), apoptosis-related proteins Caspase-3 (p17), Caspase-7 (p20), and Caspase-8 (p18), as well as necroptosis-related proteins p-MLKL, RIPK1, and RIPK3]. DSF significantly reversed the above changes induced by Ang II. Both 5z-7 and exogenous IL-1β weakened these cardioprotective effects of DSF. These results suggest that DSF may alleviate cardiac hypertrophic injury by inhibiting TAK1-mediated PANoptosis.

Key words: disulfiram; cardiac hypertrophy; TAK1; PANoptosis; H9C2 cardiomyocytes

收稿日期：　　录用日期：

通讯作者：李卫东　　E-mail:

DOI: 10.13294/j.aps.2025.0037

引用本文：

李卫东, 沈玄洋, 蒋晓璐, 文红福, 沈媛, 张美琪, 谭文涛. 双硫仑通过抑制TAK1介导的PANoptosis减轻心肌肥大损伤[J]. 生理学报 2025; 77 (2): 222-230.

LI Wei-Dong, SHEN Xuan-Yang, JIANG Xiao-Lu, WEN Hong-Fu, SHEN Yuan, ZHANG Mei-Qi, TAN Wen-Tao. Disulfiram alleviates cardiac hypertrophic injury by inhibiting TAK1-mediated PANoptosis. Acta Physiol Sin 2025; 77 (2): 222-230 (in Chinese with English abstract).