title: ER stress contributes to alpha-naphthyl isothiocyanate-induced liver injury with cholestasis in mice.
abstract: Endoplasmic reticulum (ER) stress is involved in the development of several liver diseases and tumors. This study investigated the underlying mechanisms of α-naphthyl isothiocyanate (ANIT)-induced liver injury with cholestasis in mice and found ER stress contributes to the injury. All animals were randomly divided into three groups. In the ANIT-intoxicated group, mice were intragastrically given 100mg/kg ANIT (dissolved in corn oil), while the other groups received an equal volume of vehicle as control. After 24 and 48h of ANIT administration, blood samples and liver tissues of all animals were collected for serum biochemistry and hepatic histopathological examinations to evaluate liver injuries with cholestasis. Hepatocellular apoptosis was assessed by the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. The expression of hepatic ER stress-related markers was determined by real-time PCR, immunohistochemical assay and Western blot. ANIT was found to significantly induce liver injury with cholestasis compared with control mice as evidenced by the increase of serum transaminases and total bilirubin (TBil), and histopathological changes in mice. ANIT remarkably induced hepatocellular apoptosis, upregulated the expression of caspase-9 and cytochrome c, and inhibited the gene and protein expression of proliferating cell nuclear antigen (PCNA). The gene expression of ER stress-related markers, including glucose-regulated protein 78 (GRP78), protein kinase R-like ER kinase (PERK), eukaryotic initiation factor 2α (eIF2α), inositol requiring enzyme-1α (IRE-1α) and activating transcription factor 6 (ATF6) was upregulated by ANIT in mice. ANIT also upregulated the protein expression of GRP78 and activated the phosphorylation of IRE1. These results suggested that ANIT induced liver injury with cholestasis partly due to its ability to activate the ER stress pathway.
authors.affiliation: Faculty of Pharmacy, Zhejiang Pharmaceutical College, Ningbo, 315100, China. Electronic address: 630088408@qq.com.
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Yao
authors.affiliation: Beijing Centre For Physical & Chemical Analysis, Beijing, 100050, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Li
authors.affiliation: Beijing Centre For Physical & Chemical Analysis, Beijing, 100050, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Cheng
authors.affiliation: Faculty of Pharmacy, Zhejiang Pharmaceutical College, Ningbo, 315100, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Li
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology; D053453:Caspase 9 / Q000378:metabolism; D056486:Chemical and Drug Induced Liver Injury / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D002779:Cholestasis / Q000139:chemically induced / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D045304:Cytochromes c / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D059865:Endoplasmic Reticulum Stress / Q000502:physiology*; D017953:Isocyanates; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009281:Naphthalenes
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; C000717248:Hspa5 protein, mouse; D017953:Isocyanates; D009281:Naphthalenes; C061621:alpha-naphthyl isocyanate; D045304:Cytochromes c; D053453:Caspase 9
keywords: Alpha-naphthyl isothiocyanate; Apoptosis; Cholestasis; ER stress; Liver injury