Disruption of BACH1 Protects AC16 Cardiomyocytes Against Hypoxia/Reoxygenation-Evoked Injury by Diminishing CDKN3 Transcription.

IF 3.4 3区 医学 Q2 CARDIAC & CARDIOVASCULAR SYSTEMS Cardiovascular Toxicology Pub Date : 2024-10-01 Epub Date: 2024-07-26 DOI:10.1007/s12012-024-09900-2
Yanping Li, Yi Zhou, Haifeng Pei, De Li
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Abstract

Reperfusion after myocardial infarction (MI) can lead to myocardial ischemia/reperfusion (I/R) damage. The transcription factor (TF) broad-complex, tramtrack, and bric-a-brac (BTB) and cap'n'collar (CNC) homology 1 (BACH1) is implicated in the injury. However, the downstream mechanisms of BACH1 in affecting myocardial hypoxia/reoxygenation (H/R) damage are still fully understood. AC16 cells were stimulated with H/R conditions to model cardiomyocytes under H/R. mRNA analysis was performed by quantitative real-time PCR. Protein levels were gauged by immunoblot analysis. The effect of BACH1/cyclin-dependent kinase inhibitor 3 (CDKN3) on H/R-evoked injury was assessed by measuring cell viability via Cell Counting Kit-8 (CCK-8), apoptosis (flow cytometry and caspase 3 activity), ferroptosis via Fe2+, glutathione (GSH), reactive oxygen species (ROS) and malondialdehyde (MDA) markers and inflammation cytokines interleukin-1beta (IL-1β) and tumor necrosis factor alpha (TNF-α). The BACH1/CDKN3 relationship was examined by chromatin immunoprecipitation (ChIP) experiment and luciferase assay. BACH1 was increased in MI serum and H/R-stimulated AC16 cardiomyocytes. Functionally, disruption of BACH1 mitigated H/R-evoked in vitro apoptosis, ferroptosis and inflammation of AC16 cardiomyocytes. Mechanistically, BACH1 activated CDKN3 transcription and enhanced CDKN3 protein expression in AC16 cardiomyocytes. Our rescue experiments validated that BACH1 disruption attenuated H/R-evoked AC16 cardiomyocyte apoptosis, ferroptosis and inflammation by downregulating CDKN3. Additionally, BACH1 disruption could activate the adenosine monophosphate-activated protein kinase (AMPK) signaling by downregulating CDKN3 in H/R-stimulated AC16 cardiomyocytes. Our study demonstrates that BACH1 activates CDKN3 transcription to induce H/R-evoked damage of AC16 cardiomyocytes partially via AMPK signaling.

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通过减少 CDKN3 的转录,中断 BACH1 可保护 AC16 心肌细胞免受缺氧/再氧诱发的损伤。
心肌梗塞(MI)后的再灌注可导致心肌缺血/再灌注(I/R)损伤。转录因子(TF)broad-complex、tramtrack、and bric-a-brac (BTB) and cap'n'collar (CNC) homology 1 (BACH1)与这种损伤有关。然而,BACH1影响心肌缺氧/再氧合(H/R)损伤的下游机制仍不完全清楚。用实时定量 PCR 分析 mRNA。蛋白质水平通过免疫印迹分析测定。通过细胞计数试剂盒-8(CCK-8)测量细胞活力、细胞凋亡(流式细胞术和 caspase 3 活性),评估 BACH1/细胞周期蛋白依赖性激酶抑制剂 3(CDKN3)对 H/R 诱发损伤的影响、通过Fe2+、谷胱甘肽(GSH)、活性氧(ROS)和丙二醛(MDA)标记物以及炎症细胞因子白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)来评估铁变态反应。通过染色质免疫沉淀(ChIP)实验和荧光素酶检测,研究了BACH1/CDKN3之间的关系。BACH1在MI血清和H/R刺激的AC16心肌细胞中增加。从功能上讲,破坏 BACH1 可减轻 H/R 引起的 AC16 心肌细胞体外凋亡、铁突变和炎症。从机制上讲,BACH1激活了AC16心肌细胞中CDKN3的转录并增强了CDKN3蛋白的表达。我们的抢救实验证实,BACH1中断可通过下调CDKN3减轻H/R诱发的AC16心肌细胞凋亡、铁突变和炎症。此外,在H/R刺激的AC16心肌细胞中,BACH1中断可通过下调CDKN3激活单磷酸腺苷激活蛋白激酶(AMPK)信号。我们的研究表明,BACH1可激活CDKN3转录,部分通过AMPK信号诱导AC16心肌细胞的H/R诱发损伤。
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来源期刊
Cardiovascular Toxicology
Cardiovascular Toxicology 医学-毒理学
CiteScore
6.60
自引率
3.10%
发文量
61
审稿时长
>12 weeks
期刊介绍: Cardiovascular Toxicology is the only journal dedicated to publishing contemporary issues, timely reviews, and experimental and clinical data on toxicological aspects of cardiovascular disease. CT publishes papers that will elucidate the effects, molecular mechanisms, and signaling pathways of environmental toxicants on the cardiovascular system. Also covered are the detrimental effects of new cardiovascular drugs, and cardiovascular effects of non-cardiovascular drugs, anti-cancer chemotherapy, and gene therapy. In addition, Cardiovascular Toxicology reports safety and toxicological data on new cardiovascular and non-cardiovascular drugs.
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