Identification of potential biomarkers in cholestasis and the therapeutic effect of melatonin by metabolomics, multivariate data and pathway analyses

Abstract

The present study investigated the anti-cholestatic effect of melatonin (MT) against α-naphthyl isothiocyanate (ANIT)-induced liver injury in rats and screened for potential biomarkers of cholestasis. Rats were administered ANIT by intraperitoneal injection and then sacrificed 36 h later. Serum biochemical parameters were measured and liver tissue samples were subjected to histological analysis. Active components in the serum were identified by gas chromatography-mass spectrometry, while biomarkers and biochemical pathways were identified by multivariate data analysis. The results revealed that the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, direct bilirubin, γ-glutamyl transpeptidase, and alkaline phosphatase were reduced in rats with ANIT-induced cholestasis that were treated with MT. The histological observations indicated that MT had a protective effect against ANIT-induced hepatic tissue damage. Metabolomics analysis revealed that this effect was likely to be associated with the regulation of compounds related to MT synthesis and catabolism, and amino acid metabolism, including 5-aminopentanoate, 5-methoxytryptamine, L-tryptophan, threonine, glutathione, L-methionine, and indolelactate. In addition, principal component analysis demonstrated that the levels of these metabolites differed significantly between the MT and control groups, providing further evidence that they may be responsible for the effects induced by MT. These results provide an insight into the mechanisms underlying cholestasis development and highlight potential biomarkers for disease diagnosis.

iTRAQ‑based quantitative proteomics analysis of the hepatoprotective effect of melatonin on ANIT‑induced cholestasis in rats

“…1F ). These results were consistent with those of previous studies by our group ( 11 , 12 ). For each group, 6 random fields of view were statistically evaluated for inflammatory lesions (examples in Fig.…”

“…Long-term biliary obstruction may cause liver damage, even induce biliary cirrhosis or liver fibrosis and, in severe cases, may lead to liver failure ( 25 ). In recent years, it has been indicated that oxidative stress and abnormal fatty acid metabolism may be involved in the pathogenesis of intrahepatic cholestasis ( 11 , 12 , 24 , 26 ). Previously, melatonin has been demonstrated to improve liver fibrosis and liver damage caused by various diseases through inhibiting oxidative damage ( 10 ).…”

“…There are numerous studies suggesting that melatonin may be used to treat liver disease and the hepatoprotective effects of melatonin may be associated with its antioxidant properties ( 10 ). Previous studies by our group demonstrated that the therapeutic effects of melatonin against α-naphthyl isothiocyanate (ANIT)-induced acute cholestasis are associated with the attenuation of oxidative stress ( 11 , 12 ).…”

Rodent models of cholestatic liver disease: A practical guide for translational research

“…The compound is also well known among researchers for adequately inducing cholestasis in rats and mice, 79,171 by specifically targeting bile duct epithelial cells followed by hepatocellular necrosis 137,172 . Chronic and recurrent exposure to ANIT via the diet resembles an experimental setting of chronic cholangitis, bile duct hyperplasia and peribiliary fibrosis 172‐174 . Moreover, mechanisms underlying peribiliary fibrosis in chronic ANIT models show similarities with other chronic cholestasis models, such as long‐term BDL and Mdr2 ‐/‐ mice 172 …”

Metabolomic and Lipidomic Biomarkers for Premalignant Liver Disease Diagnosis and Therapy

“…This high dose of melatonin (relative to the 4-20 mg/kg doses used in mouse melatonin studies [91,92]) produced a modest reduction in serum liver enzymes and bilirubin with a less severe liver histology. The metabolomic changes in serum due to melatonin administration were unexceptional and, in part, derived metabolically from melatonin [93]. The mechanism of ANIT-induced cholestasis continues to be investigated using metabolomic tools.…”