1. Academic Validation
  2. Hepatocytic AP-1 and STAT3 contribute to chemotaxis in alphanaphthylisothiocyanate-induced cholestatic liver injury

Hepatocytic AP-1 and STAT3 contribute to chemotaxis in alphanaphthylisothiocyanate-induced cholestatic liver injury

  • Toxicol Lett. 2022 Nov 29;373:184-193. doi: 10.1016/j.toxlet.2022.11.020.
Yishuang Luo 1 Jinyu Kang 2 Jia Luo 3 Zheng Yan 3 Shengtao Li 3 Zhuoheng Lu 3 Yufei Song 4 Xie Zhang 4 Julin Yang 5 Aiming Liu 6
Affiliations

Affiliations

  • 1 School of Medicine, Ningbo University, 315211 Ningbo, China; Ningbo Haishu District Center for Disease Control and Prevention, 315000 Ningbo, China.
  • 2 School of Medicine, Ningbo University, 315211 Ningbo, China; The Affiliated Lihuili Hospital, Ningbo University, 315000 Ningbo, China.
  • 3 School of Medicine, Ningbo University, 315211 Ningbo, China.
  • 4 The Affiliated Lihuili Hospital, Ningbo University, 315000 Ningbo, China.
  • 5 Ningbo College of Health Sciences, 315100 Ningbo, China.
  • 6 School of Medicine, Ningbo University, 315211 Ningbo, China. Electronic address: liuaiming@nbu.edu.cn.
Abstract

The development of cholestatic liver injury (CLI) involves inflammation, but the dominant pathway mediating the chemotaxis is not yet established. This work explored key signaling pathway mediating chemotaxis in CLI and the role of Kupffer cells in the inflammatory liver injury. Probe inhibitors T-5224 (100 mg/kg) for AP-1 and C188-9 (100 mg/kg) for STAT3 were used to validate key inflammatory pathways in alpha-naphthylisothiocyanate (ANIT, 100 mg/kg)-induced CLI. Two doses of GdCl3 (10 mg/kg and 40 mg/kg) were used to delete Kupffer cells and explore their role in CLI. Serum and liver samples were collected for biochemical and mechanism analysis. The liver injury in ANIT-treated mice were significantly increased supported by biochemical and histopathological changes, and neutrophils gathering around the necrotic loci. Inhibitor treatments down-regulated liver injury biomarkers except the level of total bile acid. The chemokine Ccl2 increased by 170-fold and to a less degree Cxcl2 by 45-fold after the ANIT treatment. p-c-Jun and p-STAT3 were activated in the group A but inhibited by the inhibitors in western blot analysis. The immunofluorescence results showed AP-1 not STAT3 responded to inhibitors in ANIT-induced CLI. With or without GdCl3, there was no significant difference in liver injury among the CLI groups. In necrotic loci in CLI, CXCL2 colocalized with hepatocyte biomarker Albumin, not with the F4/80 in Kupffer cells. Conclusively, AP-1 played a more critical role in the inflammation cascade than STAT3 in ANIT-induced CLI. Hepatocytes, not the Kupffer cells released chemotactic factors mediating the chemotaxis in CLI.

Keywords

AP-1; Chemotaxis; Cholestatic liver injury; Kupffer cells; STAT3.

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