NRF1诱导的mmu_circ_0001388/hsa_circ_0029470通过miR-193b-3p/TCF4/GPX4轴在缺血性急性肾损伤中赋予铁变态反应抵抗力。

IF 5.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Life sciences Pub Date : 2024-10-29 DOI:10.1016/j.lfs.2024.123190

Qiang Zheng , Dongshan Zhang , Jihong Xing

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引用次数: 0

摘要

目的：环状 RNA（circRNA）在缺血性急性肾损伤（AKI）的进展过程中至关重要。然而，mmu_circ_0001388 和 hsa_circ_0029470 的具体功能和调控途径仍不明确：方法：利用实时定量聚合酶链反应（RT-qPCR）评估 mmu_circ_0001388、hsa_circ_0029470 和 miR-139b-3p 的表达模式。通过免疫印迹鉴定了核呼吸因子 1（NRF1）、转录因子 4（TCF4）、谷胱甘肽过氧化物酶 4（GPX4）和酰基-CoA 合成酶长链家族成员 4（ACSL4）的蛋白表达。此外，还通过生物信息学预测、双荧光素酶报告（DLR）、染色质免疫沉淀（ChIP）、荧光原位杂交等多种细胞和动物实验研究了 mmu_circ_003062 和 hsa_circ_0075663 的功能和控制机制、荧光原位杂交（FISH）、流式细胞术（FCM）、苏木精和伊红（H&E）染色、二氢乙锭（DHE）、TUNEL、免疫组织化学、透射电子显微镜（TEM）和 Fe2+ 检测。主要研究结果：缺血再灌注（I/R）损伤后，最初在体外和体内观察到 NRF1 诱导 mmu_circ_0001388。随后，研究发现在波士顿大学小鼠近端肾小管（BUMPT）细胞中，敲除或过表达 mmu_circ_0001388 可分别促进或抑制 I/R 引起的铁变态反应。从机理的角度来看，研究发现 mmu_circ_0001388 可作为 miR-193b-3p 的海绵，促进 TCF4，随后增强 GPX4，从而抑制铁氧化。最后，研究表明过表达 mmu_circ_0001388 能改善小鼠 I/R 诱导的 AKI。与此同时，与 mmu_circ_0001388 同源的 hsa_circ_0029470 在人肾小管上皮细胞（HK-2）中显示出相同的控制途径：意义：NRF1/mmu_circ_0001388、hsa_circ_0029470/miR-193b-3p/TCF4/GPX4 轴在调节缺血性 AKI 诱导的铁氧化过程中起着关键作用，有望成为治疗靶点。

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NRF1-induced mmu_circ_0001388/hsa_circ_0029470 confers ferroptosis resistance in ischemic acute kidney injury via the miR-193b-3p/TCF4/GPX4 axis

Aims

Circular RNAs (circRNAs) are critical in the progression of ischemic acute kidney injury (AKI). Nevertheless, the specific functions and regulatory pathways of mmu_circ_0001388 and hsa_circ_0029470 remain elusive.

Methods

Real-time quantitative polymerase chain reaction (RT-qPCR) was utilized to assess the expression patterns of mmu_circ_0001388, hsa_circ_0029470, and miR-139b-3p. Protein expressions of nuclear respiratory factor 1 (NRF1), transcription factor 4 (TCF4), glutathione peroxidase 4 (GPX4), and Acyl-CoA synthetase long-chain family member 4 (ACSL4) were identified via immunoblotting. Furthermore, the functions and control mechanisms of mmu_circ_003062 and hsa_circ_0075663 were examined via diverse cell and animal studies, encompassing bioinformatics prediction, dual-luciferase reporter (DLR), chromatin immunoprecipitation (ChIP), fluorescence in situ hybridization (FISH), flow cytometry (FCM), hematoxylin and eosin (H&E) staining, dihydroethidium (DHE), TUNEL, immunohistochemistry, and transmission electron microscopy (TEM), and Fe²⁺ assay.

Key findings

Initially, the induction of mmu_circ_0001388 by NRF1 was observed in vitro and in vivo following ischemia/reperfusion (I/R) injury. Subsequently, knockdown or overexpression of mmu_circ_0001388 was found to either promote or inhibit ferroptosis caused by I/R in Boston University mouse proximal tubule (BUMPT) cells, respectively. From a mechanistic standpoint, mmu_circ_0001388 was found to function as a sponge for miR-193b-3p, which promoted TCF4 and subsequently enhanced GPX4, thereby suppressing ferroptosis. Finally, the overexpression of mmu_circ_0001388 was shown to ameliorate I/R-induced AKI in mice. In parallel, hsa_circ_0029470, homologous to mmu_circ_0001388, demonstrated an identical control pathway in human renal tubular epithelial (HK-2) cells.

Significance

The NRF1/mmu_circ_0001388, hsa_circ_0029470/miR-193b-3p/TCF4/GPX4 axis is pivotal in regulating ferroptosis induced by ischemic AKI and holds potential as a therapeutic target.

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来源期刊

Life sciences 医学-药学

CiteScore

12.20

自引率

1.60%

发文量

841

审稿时长

6 months

期刊介绍： Life Sciences is an international journal publishing articles that emphasize the molecular, cellular, and functional basis of therapy. The journal emphasizes the understanding of mechanism that is relevant to all aspects of human disease and translation to patients. All articles are rigorously reviewed. The Journal favors publication of full-length papers where modern scientific technologies are used to explain molecular, cellular and physiological mechanisms. Articles that merely report observations are rarely accepted. Recommendations from the Declaration of Helsinki or NIH guidelines for care and use of laboratory animals must be adhered to. Articles should be written at a level accessible to readers who are non-specialists in the topic of the article themselves, but who are interested in the research. The Journal welcomes reviews on topics of wide interest to investigators in the life sciences. We particularly encourage submission of brief, focused reviews containing high-quality artwork and require the use of mechanistic summary diagrams.