benzofurans and Cholestasis

The purpose of this study was to investigate the pharmacological effects of salvianolic acid B (SA-B) on α-naphthylisothiocyanate (ANIT)-induced cholestatic liver injury with the focus on bile acid homeostasis and anti-inflammatory pathways. Rats were randomly assigned into four groups. The control group was given normal saline (i.p.) for 7 consecutive days and on the 5th day was given the vehicle (i.g.). Model group was treated with normal saline (i.p.) for 7 days and administrated with ANIT (75 mg/kg, i.g.) on the 5th day. The SA-B groups were treated with SA-B (15 mg/kg and 30 mg/kg, i.p.) for 7 consecutive days as well as ANIT (75 mg/kg, i.g.) on the 5th day. We found that the serum levels of ALT, γ-GT, TBA, and other liver function indexes were found to be lower in the SA-B treatment groups than in the model group. SA-B also upregulated the transporters and enzymes involved in bile acid homeostasis such as Bsep, Oatp2, and Cyp3a2 in rats and BSEP, CYP3A4, and OATP2 in human cell lines. Moreover, SA-B suppressed NF-κB translocation into the nucleus, inhibited phosphorylation of p38 and JNK, and inhibited inflammation markers including IL-1β, IL-6, TGF-β, TNF-α, and COX-2 to extenuate cholestatic liver injury both in vivo and vitro. Taken together, our findings suggest that anti-cholestatic effects of SA-B may be associated with its ability to regulate NF-κB/IκB and MAPK inflammatory signaling pathways to inhibit inflammation and regulate transporters and enzymes to maintain bile acid homeostasis.

Topics: 1-Naphthylisothiocyanate; Animals; Benzofurans; Carrier Proteins; Cell Line; Cell Survival; Chemical and Drug Induced Liver Injury; Cholestasis; Cytokines; Humans; JNK Mitogen-Activated Protein Kinases; Liver; Male; Membrane Glycoproteins; NF-kappa B; NF-KappaB Inhibitor alpha; p38 Mitogen-Activated Protein Kinases; Protective Agents; Rats, Sprague-Dawley; Signal Transduction

Human toxicity screening is an important stage in the development of safe drug candidates. Hepatotoxicity is one of the major reasons for the withdrawal of drugs from the market because the liver is the major organ involved in drug metabolism, and it can generate toxic metabolites. There is a need to screen molecules for drug-induced hepatotoxicity in humans at an earlier stage. Transcriptomics is a technique widely used to screen molecules for toxicity and to unravel toxicity mechanisms. To date, the majority of such studies were performed using animals or animal cells, with concomitant difficulty in interpretation due to species differences, or in human hepatoma cell lines or cultured hepatocytes, suffering from the lack of physiological expression of enzymes and transporters and lack of nonparenchymal cells. The aim of this study was to classify known hepatotoxicants on their phenotype of toxicity in humans using gene expression profiles ex vivo in human precision-cut liver slices (PCLS). Hepatotoxicants known to induce either necrosis (n = 5) or cholestasis (n = 5) were used at concentrations inducing low (<30%) and medium (30-50%) cytotoxicity, based on ATP content. Random forest and support vector machine algorithms were used to classify hepatotoxicants using a leave-one-compound-out cross-validation method. Optimized biomarker sets were compared to derive a consensus list of markers. Classification correctly predicted the toxicity phenotype with an accuracy of 70-80%. The classification is slightly better for the low than for the medium cytotoxicity. The consensus list of markers includes endoplasmic reticulum stress genes, such as C2ORF30, DNAJB9, DNAJC12, SRP72, TMED7, and UBA5, and a sodium/bile acid cotransporter (SLC10A7). This study shows that human PCLS are a useful model to predict the phenotype of drug-induced hepatotoxicity. Additional compounds should be included to confirm the consensus list of markers, which could then be used to develop a biomarker PCR-array for hepatotoxicity screening.

Topics: Acetaminophen; Aged; Benzofurans; Bile Acids and Salts; Chloramphenicol; Chlorpromazine; Cholestasis; Colchicine; Cyclosporine; Diethylnitrosamine; Drug-Related Side Effects and Adverse Reactions; Ethinyl Estradiol; Female; Gene Expression Profiling; Hepatocytes; Humans; Liver; Male; Methyltestosterone; Middle Aged; Necrosis; Phenotype; Toxicogenetics; Young Adult

The single dose intravenous pharmacokinetics of amiodarone (50 mg/kg) were examined in rats with 72 h of biliary stasis secondary to bile duct ligation compared with paired control animals; and in rats with uranyl nitrate induced acute renal failure compared with paired control animals. Plasma and tissue levels (liver, kidney, heart, and lung) of amiodarone (1) and its N-deethyl metabolite 2 were obtained at 4 and 24 h following drug administration. Pharmacokinetic parameters were derived from plasma samples obtained over a 24-h period. Compared with controls, biliary stasis caused a decrease in the total clearance of 1 (1.74 versus 0.35 L/h/kg) and in the volume of distribution at steady state (21.1 versus 5.0 L/kg); renal failure caused a decrease in total clearance (1.67 versus 0.9 L/h/kg) and an increase in apparent elimination half-life (13.7 versus 10.1 h). Both disease processes produced significantly higher plasma levels of 1 when compared with control animals at 4 and 24 h. However, only the cholestatic animals had consistently higher tissue levels of 1 in the face of elevated plasma levels. In normal rats, no 1 or 2 was detected in the urine after a 50 mg/kg intravenous dose of 1, and less than 0.5% of the total dose of amiodarone (1) was excreted into bile by 12 h.

Topics: Amiodarone; Animals; Benzofurans; Cholestasis; Dealkylation; Glucuronates; Kidney Failure, Chronic; Kinetics; Male; Rats; Rats, Inbred Strains; Time Factors; Tissue Distribution