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

• Published in: Cellular and molecular gastroenterology and hepatology Vol. 12; no. 5; pp. 1789 - 1807
• Main Authors: Zheng, Jianxin, Zhou, Hong, Yang, Taihua, Liu, Jinchuan, Qin, Tian, Gu, Xiangqian, Wu, Ji, Zhang, Yi, Wang, Honglin, Tang, Yuanjia, Xue, Feng, Mao, Yimin, Xia, Qiang
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.2021; Elsevier
• Subjects: Acetaminophen - adverse effects; Animals; Biomarkers; Chemical and Drug Induced Liver Injury, Chronic - etiology; Chemical and Drug Induced Liver Injury, Chronic - metabolism; Chemical and Drug Induced Liver Injury, Chronic - pathology; Damage Responsive; Disease Models, Animal; Disease Susceptibility; Drug-Induced Liver Injury; Immunohistochemistry; Immunophenotyping; JNK Mitogen-Activated Protein Kinases - metabolism; MAP Kinase Signaling System; Mice; Mice, Knockout; microRNA; MicroRNAs - genetics; Models, Biological; Necrosis; Negative Feedback; Original Research; APAP; NPC; MLK3; RIP-seq; JNK; ALT; mRNA; BM; Necrosis; p-JNK; 31-KO; Damage Responsive; miRNA; GSH; WT; AST; DILI; KO; AML-12; DEG; IP; Ago; siRNA; Drug-Induced Liver Injury; MAPK; Negative Feedback; Sab; ROS; miR-31; microRNA
• ISSN: 2352-345X; 2352-345X
• DOI: 10.1016/j.jcmgh.2021.07.011

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. 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. 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. 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. [Display omitted]