NRF1 诱导的 mmu_circ_0001388/hsa_circ_0029470 通过 miR-193b-3p/TCF4/GPX4 轴赋予缺血性急性肾损伤的铁死亡抗性。
NRF1-induced mmu_circ_0001388/hsa_circ_0029470 confers ferroptosis resistance in ischemic acute kidney injury via the miR-193b-3p/TCF4/GPX4 axis.
机构信息
Department of Emergency, The First Hospital of Jilin University, Changchun 130000, Jilin, China.
Department of Emergency, The Second Xiangya Hospital of Central South University, Changsha 410000, Hunan, China; Emergency Medicine and Difficult Diseases Institute, The Second Xiangya Hospital of Central South University, Changsha 410000, Hunan, China; Department of Nephrology, The Second Xiangyi Hospital of Central South University, Changsha 410000, Hunan, China.
出版信息
Life Sci. 2024 Dec 1;358:123190. doi: 10.1016/j.lfs.2024.123190. Epub 2024 Oct 29.
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 Fe 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.
目的
环状 RNA（circRNA）在缺血性急性肾损伤（AKI）的进展中起着关键作用。然而，mmu_circ_0001388 和 hsa_circ_0029470 的具体功能和调控途径仍不清楚。
方法
实时定量聚合酶链反应（RT-qPCR）用于评估 mmu_circ_0001388、hsa_circ_0029470 和 miR-139b-3p 的表达模式。通过免疫印迹法鉴定核呼吸因子 1（NRF1）、转录因子 4（TCF4）、谷胱甘肽过氧化物酶 4（GPX4）和酰基辅酶 A 合成酶长链家族成员 4（ACSL4）的蛋白表达。此外，通过多种细胞和动物研究，包括生物信息学预测、双荧光素酶报告（DLR）、染色质免疫沉淀（ChIP）、荧光原位杂交（FISH）、流式细胞术（FCM）、苏木精和伊红（H&E）染色、二氢乙啶（DHE）、TUNEL、免疫组织化学和透射电子显微镜（TEM）以及铁测定，研究了 mmu_circ_003062 和 hsa_circ_0075663 的功能和调控机制。
主要发现
最初，在体外和体内缺血/再灌注（I/R）损伤后观察到 NRF1 诱导 mmu_circ_0001388 的诱导。随后，发现 mmu_circ_0001388 的敲低或过表达分别促进或抑制波士顿大学小鼠近端肾小管（BUMPT）细胞中由 I/R 引起的铁死亡。从机制上讲，发现 mmu_circ_0001388 作为 miR-193b-3p 的海绵起作用，miR-193b-3p 促进 TCF4，随后增强 GPX4，从而抑制铁死亡。最后，发现 mmu_circ_0001388 的过表达可改善小鼠的 I/R 诱导的 AKI。同时，与人肾近端小管上皮（HK-2）细胞中的 mmu_circ_0001388 同源的 hsa_circ_0029470 表现出相同的调控途径。
意义
NRF1/mmu_circ_0001388、hsa_circ_0029470/miR-193b-3p/TCF4/GPX4 轴在调节缺血性 AKI 诱导的铁死亡中起着关键作用，并且可能成为治疗靶点。
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