title: Alpha-naphthylisothiocyanate impairs bile acid homeostasis through AMPK-FXR pathways in rat primary hepatocytes.
abstract: Alpha-naphthylisothiocyanate (ANIT) is widely used to induce cholestasis in basic researches. Although direct damage induced by ANIT to bile duct epithelial cells has been documented in previous studies, few works investigated ANIT-induced effects on hepatocytes. Our previous study indicated that activated AMP-activated protein kinase (AMPK) inhibited farnesoid X receptor (FXR) expression and further participated in the pathogenesis of estrogen-induced cholestasis. However, whether ANIT has effects on bile acid homeostasis in hepatocytes, and the role of AMPK-FXR pathway played in these effects remain unclear. In this study, our results showed that ANIT induced intracellular bile acid accumulation without obvious cellular toxicity in sandwich cultured rat primary hepatocytes (SCRHs), accompanied with significant decreased expression of FXR and bile acid transporters. AMPK activation via ERK1/2-LKB1 pathway was critical for ANIT-induced effects on hepatocytes. Compound C, specific AMPK inhibitor, blocked ANIT-regulated gene expression, decreased bile acid accumulation and recovered bile canalicular structure both in vitro and in vivo. Furthermore, the expression of A1adenosine receptor (A1AR), a potential cholestatic target, was relatively low in hepatocytes compared with expression in rat whole livers. Consistent with these findings, DPCPX, a classic antagonist of A1AR, had no effect on ANIT-induced hepatocytes injury. In summary, our results indicate that AMPK-FXR signaling is critical for ANIT-induced toxic effects on hepatocytes, provide new insights into the pathogenesis of ANIT-induced cholestasis, and suggest AMPK-FXR pathway as a potential therapeutic target for cholestasis.
authors.affiliation: Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing, China.
authors.forename: Xiaojiaoyang
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Microbiology and Immunology, Medical College of Virginia, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Runping
authors.initials: R
authors.lastname: Liu
authors.affiliation: Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing, China.
authors.forename: Linxi
authors.initials: L
authors.lastname: Yu
authors.affiliation: Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing, China.
authors.forename: Zihang
authors.initials: Z
authors.lastname: Yuan
authors.affiliation: Department of Medical Pathomorphology, Shandong Research Academy of Traditional Chinese Medicine, Jinan, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Sun
authors.affiliation: Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing, China.
authors.forename: Hang
authors.initials: H
authors.lastname: Yang
authors.affiliation: Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing, China; Jiangsu Center for Pharmacodynamics Research and Evaluation, China Pharmaceutical University, Nanjing, China.
authors.forename: Luyong
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing, China; Jiangsu Center for Pharmacodynamics Research and Evaluation, China Pharmaceutical University, Nanjing, China. Electronic address: beaglejiang@cpu.edu.cn.
authors.forename: Zhenzhou
authors.initials: Z
authors.lastname: Jiang
mesh_terms: D015058:1-Naphthylisothiocyanate / Q000633:toxicity*; D055372:AMP-Activated Protein Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D058916:Adenosine A1 Receptor Antagonists / Q000494:pharmacology; D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism*; D001652:Bile Ducts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002779:Cholestasis / Q000139:chemically induced; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects; D006706:Homeostasis / Q000187:drug effects*; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D043682:Receptor, Adenosine A1 / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction; D014970:Xanthines / Q000494:pharmacology; D000099150:Receptor, Farnesoid X-Activated
chemical_list: D015058:1-Naphthylisothiocyanate; D055372:AMP-Activated Protein Kinases; D058916:Adenosine A1 Receptor Antagonists; D001647:Bile Acids and Salts; D043682:Receptor, Adenosine A1; D018160:Receptors, Cytoplasmic and Nuclear; D014970:Xanthines; D000099150:Receptor, Farnesoid X-Activated; C051360:1,3-dipropyl-8-cyclopentylxanthine
keywords: A(1)AR; AMPK; ANIT; Cholestasis; FXR; Hepatocyte