ZNF143 inhibits hepatocyte mitophagy and promotes non-alcoholic fatty liver disease by targeting increased lncRNA NEAT1 expression to activate ROCK2 pathway

Yujie Dong; Minjie Hu; Kewei Tan; Rongjuan Dai

doi:10.1080/15592294.2023.2239592

ZNF143 inhibits hepatocyte mitophagy and promotes non-alcoholic fatty liver disease by targeting increased lncRNA NEAT1 expression to activate ROCK2 pathway

Epigenetics. 2023 Dec;18(1):2239592. doi: 10.1080/15592294.2023.2239592.

Authors

Yujie Dong¹, Minjie Hu², Kewei Tan³, Rongjuan Dai⁴

Affiliations

¹ The First Affiliated Hospital, Department of Ultrasound Medicine, Hengyang Medical School, University of South China, Hengyang, Hunan 421001, China.
² The First Affiliated Hospital, Department of Cardiothoracic Surgery, Hengyang Medical School, University of South China, Hengyang, Hunan 421001, China.
³ The No.922 Hospital of the People Liberation Army Joint Logistics Support Force, Department of the Laboratory and Blood Transfusion, Hengyang, Hunan 421002, China.
⁴ The First Affiliated Hospital, Department of Infectious Diseases, Hengyang Medical School, University of South China, Hengyang, Hunan 421001, China.

Abstract

Background: Nonalcoholic fatty liver disease (NAFLD) is the most common hepatic disorders worldwide. The mitophagy is suggested to be repressed in NAFLD, but the mechanism remains to be elucidated.

Methods: NAFLD cell and mouse models were established by treating with free fatty acid (FFA) and feeding a high fat diet (HFD), respectively. QRT-PCR, Western blotting, or IHC measured the expression of ZNF143, lncRNA NEAT1, ROCK2, and lipid formation/mitophagy-related proteins. Cell viability and mitophagy were evaluated by MTT and immunofluorescence. The chloroform-methanol extraction method measured triglyceride and total cholesterol levels. ELISA detected ALT and AST levels. The interactions among ZNF143, lncRNA NEAT1 and SND1 were analysed by ChIP, dual-luciferase reporter, pull-down, and RIP. The lipid droplets were determined by Oil-red O and HE staining.

Results: ZNF143 and lncRNA NEAT1 were upregulated in hepatic cells treated with FFA (p < 0.01 and p < 0.001). Knockdown of ZNF143 or lncRNA NEAT1 inhibited lipid droplets formation, while promoting mitophagy (p < 0.01 and p < 0.001). ZNF143 promoted lncRNA NEAT1 transcriptional expression through binding to its promoter. LncRNA NEAT1 increased ROCK2 mRNA stability by targeting SND1. LncRNA NEAT1 or ROCK2 overexpression reversed the effect of ZNF143 or lncRNA NEAT1 knockdown on hepatic steatosis and mitophagy (p < 0.01 and p < 0.001). ZNF143 or lncRNA NEAT1 knockdown inhibited HFD-induced steatosis and promoted mitophagy in vivo (p < 0.01 and p < 0.001).

Conclusion: The upregulation of lncRNA NEAT1 caused by ZNF143 promoted NAFLD through inhibiting mitophagy via activating ROCK2 pathway by targeting SND1, providing potential targets for NAFLD therapy.

Keywords: NAFLD; ROCK2; SND1; ZNF143; lncRNA NEAT1; mitophagy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
DNA Methylation
Hepatocytes / metabolism
Liver / metabolism
Mice
MicroRNAs* / genetics
Mitophagy
Non-alcoholic Fatty Liver Disease* / genetics
RNA, Long Noncoding* / genetics
RNA, Long Noncoding* / metabolism
Trans-Activators* / metabolism

Substances

MicroRNAs
RNA, Long Noncoding
Zfp143 protein, mouse
Trans-Activators

Grants and funding

This work was supported by the Fund project of University of South China for Prevention and Control of COVID-19 [grant number 2020-22]; the Scientific Research Fund Project of Hunan Provincial Health Commission [grant number 20201983].