Department of Emergency, Third Xiangya Hospital, Central South University, Changsha 410013, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2021 Aug 28;46(8):785-792. doi: 10.11817/j.issn.1672-7347.2021.210264.
Non-alcoholic fatty liver disease has seriously affected people's health. Recent studies have found that N6-methyladenosine (m6A) methylation is involved in the lipid metabolism process of the body, but the study on the level of m6A modification in NAFLD is still not available. This study aims to explore the changes in the level of RNA m6A methylation modification in NAFLD liver tissues, and to provide experimental and theoretical basis for in-depth study on the role of RNA m6A methylation in the occurrence and development of NAFLD.
Changes in the m6A level in NAFLD liver tissues were measured by liquid chromatography-mass spectrometry (LC-MS). Total RNA was extracted from liver tissues of NAFLD patients or normal control individuals and subjected to methylated RNA immunoprecipitation (MeRIP) with microarray analysis (including 44 122 mRNAs and 12 496 lncRNAs) to determine the changes in m6A modification levels across the entire transcriptome. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to annotate the differentially modified mRNAs. Finally, 4 mRNAs and 4 lncRNAs were randomly selected to verify the microarray results by MeRIP and real-time transcription polymerase chain reaction.
A total of 176 mRNAs and 44 lncRNAs were found to be differentially m6A-modified in the NAFLD group compared with the control group. Among them, 15 mRNAs and 7 lncRNAs were hypermethylated in NAFLD, while 161 mRNAs and 37 lncRNAs were hypomethylated in NAFLD. GO and pathway analysis showed that the differentially modified mRNAs were enriched mainly in biological processes such as carboxylic acid metabolism and transcriptional regulation.
The m6A modification profile is changed in NAFLD liver tissues compared with normal liver tissues, which may functionally impact the pathophysiological progress in NAFLD.
非酒精性脂肪性肝病已严重影响人们的健康。最近的研究发现，N6-甲基腺苷（m6A）甲基化参与了机体的脂质代谢过程，但关于 NAFLD 中 m6A 修饰水平的研究仍未见报道。本研究旨在探讨非酒精性脂肪性肝病肝组织中 RNA m6A 修饰水平的变化，为深入研究 RNA m6A 甲基化在非酒精性脂肪性肝病发生发展中的作用提供实验和理论依据。
采用液相色谱-质谱联用（LC-MS）检测非酒精性脂肪性肝病肝组织中 m6A 水平的变化。提取非酒精性脂肪性肝病患者或正常对照个体肝组织中的总 RNA，采用甲基化 RNA 免疫沉淀（MeRIP）结合微阵列分析（包括 44122 个 mRNAs 和 12496 个 lncRNAs），以确定整个转录组中 m6A 修饰水平的变化。采用基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析对差异修饰的 mRNAs 进行注释。最后，随机选择 4 个 mRNAs 和 4 个 lncRNAs 通过 MeRIP 和实时转录聚合酶链反应验证微阵列结果。
与对照组相比，非酒精性脂肪性肝病组共发现 176 个 mRNAs 和 44 个 lncRNAs 存在差异 m6A 修饰。其中，15 个 mRNAs 和 7 个 lncRNAs 在非酒精性脂肪性肝病中呈高甲基化，而 161 个 mRNAs 和 37 个 lncRNAs 在非酒精性脂肪性肝病中呈低甲基化。GO 和通路分析表明，差异修饰的 mRNAs 主要富集在羧酸代谢和转录调控等生物学过程中。
与正常肝组织相比，非酒精性脂肪性肝病肝组织中的 m6A 修饰谱发生改变，可能在功能上影响非酒精性脂肪性肝病的病理生理进展。
