Mir22hg facilitates ferritinophagy-mediated ferroptosis in sepsis by recruiting the m6A reader YTHDC1 and enhancing Angptl4 mRNA stability.

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
ACS Applied Bio Materials Pub Date : 2024-08-01 Epub Date: 2024-06-06 DOI:10.1007/s10863-024-10022-1
Wenlong Deng, Liang Zhong, Shupei Ye, Jiajing Luo, Guobin Ren, Junhao Huang, Xiaolei Zhuang
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Abstract

Background: Ferritinophagy-mediated ferroptosis plays a crucial role in fighting pathogen aggression. The long non-coding RNA Mir22hg is involved in the regulation of ferroptosis and aberrantly overexpression in lipopolysaccharide (LPS)-induced sepsis mice, but whether it regulates sepsis through ferritinophagy-mediated ferroptosis is unclear.

Methods: Mir22hg was screened by bioinformatics analysis. Ferroptosis was assessed by assaying malondialdehyde (MDA), reactive oxygen species (ROS), and Fe2+ levels, glutathione (GSH) activity, as well as ferroptosis-related proteins GPX4 and SLC3A2 by using matched kits and performing western blot. Ferritinophagy was assessed by Lyso tracker staining and FerroOrange staining, immunofluorescence analysis of Ferritin and LC-3, and western blot analysis of LC-3II/I, p62, FTH1, and NCOA4. The bind of YTH domain containing 1 (YTHDC1) to Mir22hg or angiopoietin-like-4 (Angptl4) was verified by RNA pull-down and/or immunoprecipitation (RIP) assays.

Results: Mir22hg silencing lightened ferroptosis and ferritinophagy in LPS-induced MLE-12 cells and sepsis mouse models, as presented by the downregulated MDA, ROS, Fe2+, NCOA4, and SLC3A2 levels, upregulated GPX4, GSH, and FTH1 levels, along with a decrease in autophagy. Mir22hg could bind to the m6A reader YTHDC1 without affecting its expression. Mechanistically, Mir22hg enhanced Angptl4 mRNA stability through recruiting the m6A reader YTHDC1. Furthermore, Angptl4 overexpression partly overturned Mir22hg inhibition-mediated effects on ferroptosis and ferritinophagy in LPS-induced MLE-12 cells.

Conclusion: Mir22hg contributed to in ferritinophagy-mediated ferroptosis in sepsis via recruiting the m6A reader YTHDC1 and strengthening Angptl4 mRNA stability, highlighting that Mir22hg may be a potential target for sepsis treatment based on ferroptosis.

Abstract Image

Mir22hg 通过招募 m6A 阅读器 YTHDC1 和增强 Angptl4 mRNA 的稳定性,促进败血症中由铁蛋白吞噬介导的铁蛋白沉积。
背景:铁蛋白吞噬介导的铁蛋白沉积在对抗病原体侵袭中起着至关重要的作用。长非编码 RNA Mir22hg 参与调控铁吞蛋白,并在脂多糖(LPS)诱导的败血症小鼠中异常过表达,但其是否通过噬铁蛋白介导的铁吞蛋白调控败血症尚不清楚:方法:通过生物信息学分析筛选 Mir22hg。方法:通过生物信息学分析筛选出 Mir22hg,使用匹配的试剂盒并进行 Western 印迹,通过检测丙二醛(MDA)、活性氧(ROS)和 Fe2+ 水平、谷胱甘肽(GSH)活性以及铁氧化相关蛋白 GPX4 和 SLC3A2 来评估铁氧化。通过Lyso tracker染色和FerroOrange染色、铁蛋白和LC-3的免疫荧光分析以及LC-3II/I、p62、FTH1和NCOA4的Western印迹分析来评估铁蛋白吞噬作用。通过RNA牵引和/或免疫沉淀(RIP)实验验证了含YTH结构域的1(YTHDC1)与Mir22hg或血管生成素样-4(Angptl4)的结合:结果:在LPS诱导的MLE-12细胞和脓毒症小鼠模型中,沉默Mir22hg可减轻铁变态反应和铁蛋白吞噬,表现为MDA、ROS、Fe2+、NCOA4和SLC3A2水平下调,GPX4、GSH和FTH1水平上调,自噬减少。Mir22hg 能与 m6A 阅读器 YTHDC1 结合而不影响其表达。从机制上讲,Mir22hg通过招募m6A阅读器YTHDC1增强了Angptl4 mRNA的稳定性。此外,在LPS诱导的MLE-12细胞中,Angptl4的过表达部分推翻了Mir22hg抑制介导的对铁蛋白吞噬和铁蛋白吞噬的影响:结论:Mir22hg通过招募m6A阅读器YTHDC1和加强Angptl4 mRNA的稳定性,促进了脓毒症中铁蛋白吞噬介导的铁蛋白沉积,突出表明Mir22hg可能是基于铁蛋白沉积治疗脓毒症的潜在靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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