1. Academic Validation
  2. Protective Role of microRNA-31 in Acetaminophen-Induced Liver Injury: A Negative Regulator of c-Jun N-Terminal Kinase (JNK) Signaling Pathway

Protective Role of microRNA-31 in Acetaminophen-Induced Liver Injury: A Negative Regulator of c-Jun N-Terminal Kinase (JNK) Signaling Pathway

  • Cell Mol Gastroenterol Hepatol. 2021;12(5):1789-1807. doi: 10.1016/j.jcmgh.2021.07.011.
Jianxin Zheng 1 Hong Zhou 2 Taihua Yang 3 Jinchuan Liu 3 Tian Qin 3 Xiangqian Gu 3 Ji Wu 3 Yi Zhang 4 Honglin Wang 5 Yuanjia Tang 6 Feng Xue 7 Yimin Mao 8 Qiang Xia 3
Affiliations

Affiliations

  • 1 Department of Liver Surgery and Liver Transplantation Center, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China; Department of Urology, Xiamen Branch, Zhongshan Hospital, Fudan University, Xiamen, China.
  • 2 Center for Microbiota and Immunological Diseases, Shanghai General Hospital, Shanghai Institute of Immunology, Shanghai Jiaotong University School of Medicine, Shanghai, China.
  • 3 Department of Liver Surgery and Liver Transplantation Center, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
  • 4 ABLife Institute of BioBigData, East Lake High-Tech Development Zone, Wuhan, China.
  • 5 Center for Microbiota and Immunological Diseases, Shanghai General Hospital, Shanghai Institute of Immunology, Shanghai Jiaotong University School of Medicine, Shanghai, China. Electronic address: [email protected].
  • 6 Shanghai Institute of Rheumatology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China. Electronic address: [email protected].
  • 7 Department of Liver Surgery and Liver Transplantation Center, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China. Electronic address: [email protected].
  • 8 Division of Gastroenterology and Hepatology, Shanghai Institute of Digestive Disease, Shanghai Jiaotong University School of Medicine, Shanghai, China.
Abstract

Background & aims: Sustained c-Jun N-terminal kinase (JNK) activation plays a major role in drug-induced liver injury (DILI). Stress-responsive microRNA-31 (miR-31) has been implicated in regulating different cellular damage, and JNK activation could induce miR-31 expression. However, the regulatory role of miR-31 in DILI has not been studied previously. We aimed to investigate whether miR-31 could ameliorate DILI and ascertain potential molecular mechanism.

Methods: miR-31 gene knockout (31-KO) and wild-type C57BL/6J mice were used to construct an acetaminophen (APAP)-induced DILI model. Primary mouse hepatocytes, as well as alpha mouse liver 12 (AML-12) cell lines, were used for in vitro experiments. Argonaute 2-associated RNA immunoprecipitation combined with high-throughput sequencing were performed to identify specific targets of miR-31.

Results: 31-KO mice showed a higher mortality rate, liver transaminase levels, and hepatic necrosis compared with those in wild-type mice after APAP-induced hepatotoxicity. The protective role of miR-31 on hepatocytes has been analyzed via constructing bone marrow chimeric mice. Mechanistically, we found that hepatic JNK phosphorylation increased significantly in 31-KO mice. This caused mitochondrial phosphorylated Src (p-Src) inactivation and more Reactive Oxygen Species production, which directly amplifies hepatocyte necrotic cell death, while administration of JNK-specific inhibitor SP600125 could abrogate the differences. Moreover, bioinformatics analysis of RNA immunoprecipitation combined with high-throughput sequencing identified that guanosine triphosphatase, cell division cycle protein 42 (Cdc42), the upstream molecule of JNK signaling, was the specific target of miR-31 and could form a miR-31/Cdc42/phosphorylated mixed-lineage kinase 3 (p-MLK3) negative feedback loop to restrict JNK overactivation. Clinically, both miR-31 and phosphorylated JNK (p-JNK) were highly increased in liver tissues of DILI patients with different etiologies.

Conclusions: miR-31 can down-regulate Cdc42 to restrict overactivation of Reactive Oxygen Species/JNK/mitochondria necrotic death loop in hepatocytes of APAP-induced DILI, which might provide a new therapeutic target for alleviating JNK overactivation-based liver injury.

Keywords

Damage Responsive; Drug-Induced Liver Injury; Necrosis; Negative Feedback; microRNA.

Figures
Products