LINC00894 Regulates Cerebral Ischemia/Reperfusion Injury by Stabilizing EIF5 and Facilitating ATF4-Mediated Induction of FGF21 and ACOD1 Expression

IF 3.7 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Neurochemical Research Pub Date : 2024-07-26 DOI:10.1007/s11064-024-04213-w

Yifei Chen, Hengxiang Cui, Zhuanzhuan Han, Lei Xu, Lin Wang, Yuefei Zhang, Lijun Liu

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Abstract

The non-coding RNA LINC00894 modulates tumor proliferation and drug resistance. However, its role in brain is still unclear. Using RNA-pull down combined with mass spectrometry and RNA binding protein immunoprecipitation, EIF5 was identified to interact with LINC00894. Furthermore, LINC00894 knockdown decreased EIF5 protein expression, whereas LINC00894 overexpression increased EIF5 protein expression in SH-SY5Y and BE(2)-M17 (M17) neuroblastoma cells. Additionally, LINC00894 affected the ubiquitination modification of EIF5. Adeno-associated virus (AAV) mediated LINC00894 overexpression in the brain inhibited the expression of activated Caspase-3, while increased EIF5 protein level in rats and mice subjected to transient middle cerebral artery occlusion reperfusion (MCAO/R). Meanwhile, LINC00894 knockdown increased the number of apoptotic cells and expression of activated Caspase-3, and its overexpression decreased them in the oxygen–glucose deprivation and reoxygenation (OGD/R) in vitro models. Further, LINC00894 was revealed to regulated ATF4 protein expression in condition of OGD/R and normoxia. LINC00894 knockdown also decreased the expression of glutamate-cysteine ligase catalytic subunit (GCLC) and ATF4, downregulated glutathione (GSH), and the ratio of GSH to oxidized GSH (GSH: GSSG) in vitro. By using RNA-seq combined with qRT-PCR and immunoblot, we identified that fibroblast growth factor 21 (FGF21) and aconitate decarboxylase 1 (ACOD1), as the ATF4 target genes were regulated by LINC00894 in the MCAO/R model. Finally, we revealed that ATF4 transcriptionally regulated FGF21 and ACOD1 expression; ectopic overexpression of FGF21 or ACOD1 in LINC00894 knockdown cells decreased activated Caspase-3 expression in the OGD/R model. Our results demonstrated that LINC00894 regulated cerebral ischemia injury by stabilizing EIF5 and facilitating EIF5-ATF4-dependent induction of FGF21 and ACOD1.

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LINC00894 通过稳定 EIF5 和促进 ATF4 介导的 FGF21 和 ACOD1 表达调控脑缺血再灌注损伤

非编码 RNA LINC00894 可调节肿瘤增殖和耐药性。然而，它在大脑中的作用仍不清楚。利用 RNA 拉低结合质谱法和 RNA 结合蛋白免疫沉淀法，确定了 EIF5 与 LINC00894 的相互作用。此外，在SH-SY5Y和BE（2）-M17（M17）神经母细胞瘤细胞中，LINC00894敲除会降低EIF5蛋白的表达，而LINC00894过表达则会增加EIF5蛋白的表达。此外，LINC00894 还影响了 EIF5 的泛素化修饰。腺相关病毒（AAV）介导的LINC00894过表达抑制了活化Caspase-3的表达，同时提高了一过性大脑中动脉闭塞再灌注（MCAO/R）大鼠和小鼠的EIF5蛋白水平。同时，在氧-葡萄糖剥夺和再氧合（OGD/R）体外模型中，LINC00894敲除会增加凋亡细胞的数量和活化Caspase-3的表达，而过表达则会降低它们。此外，LINC00894 还能调节 ATF4 蛋白在 OGD/R 和常氧条件下的表达。LINC00894 基因敲除还降低了谷氨酸-半胱氨酸连接酶催化亚基（GCLC）和 ATF4 的表达，下调了谷胱甘肽（GSH）以及体外 GSH 与氧化 GSH 的比率（GSH：GSSG）。通过RNA-seq结合qRT-PCR和免疫印迹，我们发现成纤维细胞生长因子21（FGF21）和醋酸脱羧酶1（ACOD1）作为ATF4靶基因在MCAO/R模型中受到LINC00894的调控。最后，我们发现 ATF4 可转录调控 FGF21 和 ACOD1 的表达；在 OGD/R 模型中，在 LINC00894 敲除的细胞中异位过表达 FGF21 或 ACOD1 可降低活化的 Caspase-3 的表达。我们的研究结果表明，LINC00894通过稳定EIF5和促进EIF5-ATF4依赖性诱导FGF21和ACOD1来调节脑缺血损伤。

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来源期刊

Neurochemical Research 医学-神经科学

CiteScore

7.70

自引率

2.30%

发文量

320

审稿时长

6 months

期刊介绍： Neurochemical Research is devoted to the rapid publication of studies that use neurochemical methodology in research on nervous system structure and function. The journal publishes original reports of experimental and clinical research results, perceptive reviews of significant problem areas in the neurosciences, brief comments of a methodological or interpretive nature, and research summaries conducted by leading scientists whose works are not readily available in English.