通过 RNA-seq 分析探讨 ADAR1 缺失诱导的非酒精性脂肪性肝病中与铁死亡相关的基因。

Exploration and Validation of Ferroptosis-Associated Genes in ADAR1 Deletion-Induced NAFLD through RNA-seq Analysis.

机构信息

Henan Key Laboratory of Helicobacter Pylori, Microbiota and Gastrointestinal Cancer, Marshall Medical Research Center, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China; Academy of Medical Sciences, Zhengzhou University, Zhengzhou, Henan, China; Department of Gastroenterology, the Fifth Affiliated Hospital, Zhengzhou University, Zhengzhou, Henan, China.

Henan Key Laboratory of Helicobacter Pylori, Microbiota and Gastrointestinal Cancer, Marshall Medical Research Center, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.

出版信息

Int Immunopharmacol. 2024 Jun 15;134:112177. doi: 10.1016/j.intimp.2024.112177. Epub 2024 May 1.

DOI:10.1016/j.intimp.2024.112177

PMID:38696908

Abstract

BACKGROUND

Ferroptosis, characterized by excessive iron ions and lipid peroxides accumulation, contributes to Nonalcoholic Fatty Liver Disease (NAFLD) development. The role of ADAR1, crucial for lipid metabolism and immune regulation, in ferroptosis-related NAFLD remains unexplored.

METHODS

In this study, we analyzed the expression of ADAR1 in NAFLD patients using the GSE66676 database. Subsequently, We investigated the effects of ADAR1 knockdown on mitochondrial membrane potential (MMP), Fe2+ levels, oxidation products, and ferroptosis in NAFLD cells through in vitro and in vivo experiments. Additionally, RNA-seq analysis was performed following ADAR1 depletion in an NAFLD cell model. Overlapping and ferroptosis-related genes were identified using a Venn diagram, while Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were conducted as well. Furthermore, a protein-protein interaction (PPI) network was constructed to identify hub genes associated with ferroptosis.

RESULTS

We found the expression level of ADAR1 was downregulated in NAFLD patients and 22 ferroptosis-associated genes were differentially expressed in a NAFLD cell model upon ADAR1 knockdown. Based on PPI network, we identified NOS2, PTGS2, NOX4, ALB, IL6, and CCL5 as the central genes related to ferroptosis. ADAR1 deletion-related NAFLD was found to be involved in the ferroptosis signaling pathway. NOS2, PTGS2, ALB, and IL6 can serve as potential biomarkers. These findings offer new insights and expanded targets for NAFLD prevention and treatment.

CONCLUSION

These findings provide new strategies and potential targets for preventing and treating NAFLD. NOS2, PTGS2, ALB, and IL6 may serve as biomarkers for ADAR1 deletion-related NAFLD, which could help for developing its new diagnostic and therapeutic strategies.

摘要

背景

铁死亡是一种由铁离子和脂质过氧化物积累引起的疾病，与非酒精性脂肪性肝病（NAFLD）的发展有关。ADAR1 在脂质代谢和免疫调节中起着至关重要的作用，但它在铁死亡相关的 NAFLD 中的作用仍未被探索。

方法

本研究通过 GSE66676 数据库分析了 ADAR1 在 NAFLD 患者中的表达情况。随后，通过体外和体内实验研究了 ADAR1 敲低对 NAFLD 细胞中线粒体膜电位（MMP）、Fe2+水平、氧化产物和铁死亡的影响。此外，在 NAFLD 细胞模型中，通过 ADAR1 敲除进行了 RNA-seq 分析。通过 Venn 图识别重叠和铁死亡相关基因，同时进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析。此外，构建了一个蛋白质-蛋白质相互作用（PPI）网络，以识别与铁死亡相关的枢纽基因。

结果

我们发现 ADAR1 的表达水平在 NAFLD 患者中下调，并且在 ADAR1 敲低的 NAFLD 细胞模型中，有 22 个铁死亡相关基因表达差异。基于 PPI 网络，我们确定 NOS2、PTGS2、NOX4、ALB、IL6 和 CCL5 为与铁死亡相关的核心基因。ADAR1 缺失相关的 NAFLD 被发现与铁死亡信号通路有关。NOS2、PTGS2、ALB 和 IL6 可以作为潜在的生物标志物。这些发现为 NAFLD 的预防和治疗提供了新的思路和扩展的靶点。