Pediatric Department, The Second Affiliated Hospital of Nantong University, Nantong City, Jiangsu Province 226006, China.
Affiliated Maternity and Child Health Care Hospital of Nantong University.
Crit Rev Eukaryot Gene Expr. 2024;34(8):47-57. doi: 10.1615/CritRevEukaryotGeneExpr.2024054011.
FTO alpha-ketoglutarate dependent dioxygenase (FTO) is aberrantly expressed in brain disorders. However, the roles of FTO in neonatal hypoxic-ischemic brain injury (HIE) are still unclear. This study aims to investigate the potential of FTO in neonatal HIE. Oxygen-glucose deprivation (OGD) was used to establish HIE in vitro. mRNA levels were detected by real-time reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). Protein expression was detected by Western blot. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), ferrous iron (Fe2+) and glutathione (GSH) was detected by specific kit. m6A sites were analyzed using SRAMP and further verify by methylated RNA immunoprecipitation (MeRIP) assay. Cell proliferation was determined by Cell Counting Kit-8 (CCK-8) assay. Cell death was determined by propidium iodide (PI) staining. FTO was downregulated in patients with neonatal HIE and OGD-treated neurons. Moreover, FTO mRNA expression was decreased in ferroptosis inducer, especially ferric ammonium citrate (FAC). However, overexpression of FTO inhibited the ferroptosis of neurons. Moreover, FTO-mediated N6-methyladenosine (m6A) modification of ferritin heavy chain 1 (FTH1) suppressed its mRNA expression and stability, inhibiting its protein expression. However, overexpression of FTH1 abrogated the effects of FTO and promoted the ferroptosis of neurons. In summary, FTO functions as a protective role in neonatal HIE via inhibiting FTH1 signaling. Thence, targeting may be a promising strategy for FTO neonatal HIE.
脂肪量和肥胖相关基因（FTO）α-酮戊二酸依赖性双加氧酶（FTO）在脑部疾病中异常表达。然而，FTO 在新生儿缺氧缺血性脑损伤（HIE）中的作用仍不清楚。本研究旨在探讨 FTO 在新生儿 HIE 中的潜在作用。使用氧葡萄糖剥夺（OGD）建立体外 HIE 模型。实时逆转录定量聚合酶链反应（RT-qPCR）检测 mRNA 水平。Western blot 检测蛋白表达。采用特定试剂盒检测丙二醛（MDA）、超氧化物歧化酶（SOD）、亚铁离子（Fe2+）和谷胱甘肽（GSH）的水平。使用 SRAMP 分析 m6A 位点，并通过甲基化 RNA 免疫沉淀（MeRIP）检测进一步验证。通过细胞计数试剂盒-8（CCK-8）检测细胞增殖。通过碘化丙啶（PI）染色检测细胞死亡。新生儿 HIE 患者和 OGD 处理神经元中 FTO 下调。此外，铁死亡诱导剂，特别是柠檬酸铁铵（FAC），FTO mRNA 表达降低。然而，FTO 的过表达抑制了神经元的铁死亡。此外，FTO 介导的铁蛋白重链 1（FTH1）的 N6-甲基腺苷（m6A）修饰抑制其 mRNA 表达和稳定性，抑制其蛋白表达。然而，FTH1 的过表达消除了 FTO 的作用并促进了神经元的铁死亡。总之，FTO 通过抑制 FTH1 信号在新生儿 HIE 中发挥保护作用。因此，靶向可能是治疗 FTO 新生儿 HIE 的有前途的策略。
