METTL14 Aggravates Sepsis-Induced Acute Kidney Injury by Promoting Ferroptosis Through m6A Modification of BMAL1

IF 2.5 4区医学 Q2 Medicine

Clinical and Experimental Pharmacology and Physiology Pub Date : 2026-03-08 DOI:10.1111/1440-1681.70107

Yishu Wang, Yang Li, Na Wu, Xinyue Xu, Xiaoxuan Fan, Haifeng Wang

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Abstract

Background

Acute kidney injury (AKI) is a common and severe complication of sepsis and represents an independent risk factor for mortality in septic patients. Despite its clinical significance, the mechanisms of sepsis-induced AKI (Sepsis-AKI) remain incompletely understood. This study investigates the role of methyltransferase like 14 (METTL14)-mediated m6A modification in regulating brain and muscle ARNT-like protein-1 (BMAL1) stability and its effect on tubular epithelial cell injury and ferroptosis.

Methods

Human renal proximal tubular epithelial (HK-2) cells were treated with lipopolysaccharide (LPS) to establish an in vitro model of Sepsis-AKI. Cell proliferation and viability were assessed using EdU and CCK-8 assays; apoptosis was evaluated by TUNEL staining, and inflammatory cytokines Interleukin-6 (IL-6) and IL-1β were measured by ELISA. Ferroptosis indices were detected using corresponding kits. RT-qPCR and Western blotting were used to detect mRNA and protein expression. MeRIP and RIP assays were used to evaluate BMAL1 m6A modification and RNA-protein interaction. The stability of BMAL1 mRNA was determined using an Actinomycin D chase assay. A Sepsis-AKI model was established to examine the effect of METTL14 silencing on renal injury.

Results

BMAL1 overexpression significantly alleviated LPS-induced apoptosis, inflammatory responses, and ferroptosis in HK-2 cells. Furthermore, METTL14 silencing reduced BMAL1 m6A modification, stabilized BMAL1 mRNA, and consequently improved HK-2 cell injury. In addition, YTHN6-methyladenosine RNA binding protein 1 (YTHDF1) was identified as the critical m6A reader mediating BMAL1 mRNA degradation. Consistently, in vivo experiments demonstrated that METTL14 knockdown mitigated Sepsis-AKI and ferroptosis in mice.

Conclusion

METTL14 enhanced BMAL1 m6A modification and promoted YTHDF1-mediated BMAL1 degradation, thereby facilitating ferroptosis and aggravating Sepsis-AKI.

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METTL14通过m6A修饰BMAL1促进铁下沉加重脓毒症诱导的急性肾损伤

背景：急性肾损伤（AKI）是脓毒症常见且严重的并发症，是脓毒症患者死亡的独立危险因素。尽管具有临床意义，但脓毒症诱发AKI （Sepsis-AKI）的机制仍不完全清楚。本研究探讨甲基转移酶样14 （METTL14）介导的m6A修饰在调节脑和肌肉art样蛋白-1 （BMAL1）稳定性中的作用及其对小管上皮细胞损伤和铁上吊的影响。方法：采用脂多糖（LPS）处理人肾近端小管上皮细胞（HK-2），建立脓毒症- aki体外模型。用EdU和CCK-8测定细胞增殖和活力；TUNEL染色检测细胞凋亡，ELISA检测炎症因子白介素-6 （IL-6）、IL-1β水平。采用相应试剂盒检测下垂铁指数。RT-qPCR和Western blotting检测mRNA和蛋白的表达。MeRIP和RIP检测BMAL1 m6A修饰和rna -蛋白相互作用。采用放线菌素D追踪法测定BMAL1 mRNA的稳定性。建立脓毒症- aki模型，观察METTL14沉默对肾损伤的影响。结果：BMAL1过表达可显著减轻lps诱导的HK-2细胞凋亡、炎症反应和铁下垂。此外，METTL14沉默减少了BMAL1 m6A修饰，稳定了BMAL1 mRNA，从而改善了HK-2细胞损伤。此外，ythn6 -甲基腺苷RNA结合蛋白1 （YTHDF1）被鉴定为介导BMAL1 mRNA降解的关键m6A读取器。与此一致的是，体内实验表明，METTL14敲低可减轻小鼠脓毒症- aki和铁下垂。结论：METTL14增强BMAL1 m6A修饰，促进ythdf1介导的BMAL1降解，从而促进铁下沉，加重脓毒症- aki。

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来源期刊

Clinical and Experimental Pharmacology and Physiology 医学-生理学

CiteScore

6.20

自引率

0.00%

发文量

128

审稿时长

6 months

期刊介绍： Clinical and Experimental Pharmacology and Physiology is an international journal founded in 1974 by Mike Rand, Austin Doyle, John Coghlan and Paul Korner. Our focus is new frontiers in physiology and pharmacology, emphasizing the translation of basic research to clinical practice. We publish original articles, invited reviews and our exciting, cutting-edge Frontiers-in-Research series’.