Dexamethasone counteracts hepatic inflammation and oxidative stress in cholestatic rats via CAR activation

Glucocorticoids (GCs) are currently used for the therapeutic management of cholestatic diseases, but their use and molecular mechanism remain controversial. The aims of this study were 1) to assess the therapeutic effect of a 2-week treatment with the GC dexamethasone on hepatic damage in bile duct-ligated rats; 2) to investigate its effect on the activation of the nuclear receptors (NRs) pregnane X receptor (PXR), constitutive androstane receptor (CAR) and GC receptor (GR), and NF-kB, as well as on oxidative stress and bile acid (BA) hepatic composition. Cholestasis was induced by ligation of bile duct (BDL animals) in 16 male Wistar-Kyoto rats, and eight of them were daily treated by oral gavage with 0.125 mg/ml/kg DEX for 14 days. Eight Sham-operated rats were used as controls. Severity of cholestasis was assessed histologically and on plasma biochemical parameters. The nuclear expression of NF-kB (p65), GR, PXR and CAR was measured in hepatic tissue by Western Blot. Oxidative stress was evaluated by measuring malondialdehyde, carbonylated proteins, GHS and ROS content in rat livers. LC-MS was used to measure the plasma and liver concentration of 7 BAs. Histological findings and a significant drop in several markers of inflammation (p65 nuclear translocation, mRNA expressions of TNF-α, IL-1β, IL-6) showed that DEX treatment reversed cholestasis-induced inflammation, and similar results have been obtained with oxidative stress markers. The nuclear expression of p65 and CAR were inversely correlated, with the latter increasing significantly after DEX treatment (p<0.01 vs vehicle). Hepatic BA levels tended to drop in the untreated cholestatic rats, whereas they were similar to those of healthy rats in DEX-treated animals. Plasma BAs decreased significantly in DEX-treated animals with respect to untreated cholestatic rats. In conclusion, DEX reduces inflammation and oxidative stress in BDL rats, and probably CAR is responsible for this effect. Therefore, this NR represents a promising pharmacological target for managing cholestatic and inflammatory liver diseases.

[1]  M. Carrara,et al.  Western Diet-Induced Metabolic Alterations Affect Circulating Markers of Liver Function before the Development of Steatosis , 2019, Nutrients.

[2]  F. Farinati,et al.  Focus on histological abnormalities of intrahepatic vasculature in chronic viral hepatitis , 2018, Liver international : official journal of the International Association for the Study of the Liver.

[3]  D. Petersen,et al.  Dysregulation of antioxidant responses in patients diagnosed with concomitant Primary Sclerosing Cholangitis/Inflammatory Bowel Disease. , 2018, Experimental and molecular pathology.

[4]  Roberta Lazzari,et al.  Pregnane X receptor and constitutive androstane receptor modulate differently CYP3A-mediated metabolism in early- and late-stage cholestasis , 2017, World journal of gastroenterology.

[5]  Luyong Zhang,et al.  Quantitative profiling of 19 bile acids in rat plasma, liver, bile and different intestinal section contents to investigate bile acid homeostasis and the application of temporal variation of endogenous bile acids , 2017, The Journal of Steroid Biochemistry and Molecular Biology.

[6]  Yoon Jae Kim,et al.  Ursodeoxycholic acid inhibits the proliferation of colon cancer cells by regulating oxidative stress and cancer stem-like cell growth , 2017, PloS one.

[7]  Q. Kan,et al.  Alterations of Histone Modifications Contribute to Pregnane X Receptor-Mediated Induction of CYP3A4 by Rifampicin , 2017, Molecular Pharmacology.

[8]  R. Heidari,et al.  Sulfasalazine induces mitochondrial dysfunction and renal injury , 2017, Renal failure.

[9]  L. Facci,et al.  An intracellular adrenomedullin system reduces IL-6 release via a NF-kB-mediated, cAMP-independent transcriptional mechanism in rat thymic epithelial cells. , 2016, Cytokine.

[10]  T. Matsuura,et al.  Oxidative stress markers, secondary bile acids and sulfated bile acids classify the clinical liver injury type: Promising diagnostic biomarkers for cholestasis. , 2016, Chemico-biological interactions.

[11]  S. Strom,et al.  Addition of Dexamethasone Alters the Bile Acid Composition by Inducing CYP8B1 in Primary Cultures of Human Hepatocytes. , 2016, Journal of clinical and experimental hepatology.

[12]  P. Palatini,et al.  Pharmacokinetic drug interactions in liver disease: An update. , 2016, World journal of gastroenterology.

[13]  Liu Jc,et al.  Oxidative stress markers in intrahepatic cholestasis of pregnancy: a prospective controlled study. , 2015 .

[14]  E. Elias,et al.  Rifampicin in the treatment of severe intrahepatic cholestasis of pregnancy. , 2015, European journal of obstetrics, gynecology, and reproductive biology.

[15]  Sabine Weiskirchen,et al.  Bile Duct Ligation in Mice: Induction of Inflammatory Liver Injury and Fibrosis by Obstructive Cholestasis , 2015, Journal of visualized experiments : JoVE.

[16]  G. Albertin,et al.  Differential Effect of Liver Cirrhosis on the Pregnane X Receptor–Mediated Induction of CYP3A1 and 3A2 in the Rat , 2014, Drug Metabolism and Disposition.

[17]  Fan Wang,et al.  Combination Therapy of Ursodeoxycholic Acid and Corticosteroids for Primary Biliary Cirrhosis with Features of Autoimmune Hepatitis: A Meta-Analysis , 2013, Gastroenterology research and practice.

[18]  R. Orlando,et al.  Severe Liver Cirrhosis Markedly Reduces AhR-Mediated Induction of Cytochrome P450 in Rats by Decreasing the Transcription of Target Genes , 2013, PloS one.

[19]  J. Boyer,et al.  Molecular pathogenesis of cholestasis. , 2012, The New England journal of medicine.

[20]  J. Turnay,et al.  Deoxycholic and chenodeoxycholic bile acids induce apoptosis via oxidative stress in human colon adenocarcinoma cells , 2011, Apoptosis.

[21]  S. Fuchs,et al.  Blocking of IL-6 suppresses experimental autoimmune myasthenia gravis. , 2011, Journal of autoimmunity.

[22]  A. Ramos,et al.  Oxidative status, in vitro iron-induced lipid oxidation and superoxide dismutase, catalase and glutathione peroxidase activities in rhea meat. , 2010, Meat science.

[23]  T. Hervig,et al.  A novel HPLC method for the measurement of thiobarbituric acid reactive substances (TBARS). A comparison with a commercially available kit. , 2006, Clinical biochemistry.

[24]  M. Pfaffl,et al.  Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR , 2006, Biotechnology Letters.

[25]  J. Pascussi,et al.  Is nuclear factor kappa-B the missing link between inflammation, cancer and alteration in hepatic drug metabolism in patients with cancer? , 2006, European journal of cancer.

[26]  P. Gervasi,et al.  CAR and PXR expression and inducibility of CYP2B and CYP3A activities in rat and rabbit lungs. , 2005, Life sciences.

[27]  Micheline Piquette-Miller,et al.  The Involvement of the Pregnane X Receptor in Hepatic Gene Regulation during Inflammation in Mice , 2005, Journal of Pharmacology and Experimental Therapeutics.

[28]  W. Lamers,et al.  Mechanisms of glucocorticoid signalling. , 2004, Biochimica et biophysica acta.

[29]  K. Brouwer,et al.  Effect of dexamethasone treatment on the expression and function of transport proteins in sandwich-cultured rat hepatocytes. , 2004, Drug metabolism and disposition: the biological fate of chemicals.

[30]  P. Gatellier,et al.  Lipid and protein oxidation in vitro, and antioxidant potential in meat from Charolais cows finished on pasture or mixed diet. , 2004, Meat science.

[31]  K. Feingold,et al.  Reduction in cytochrome P-450 enzyme expression is associated with repression of CAR (constitutive androstane receptor) and PXR (pregnane X receptor) in mouse liver during the acute phase response. , 2002, Biochemical and biophysical research communications.

[32]  I. Adcock,et al.  p65-activated Histone Acetyltransferase Activity Is Repressed by Glucocorticoids , 2001, The Journal of Biological Chemistry.

[33]  I. Adcock,et al.  Glucocorticoid-regulated transcription factors. , 2001, Pulmonary pharmacology & therapeutics.

[34]  J. Pascussi,et al.  Dexamethasone enhances constitutive androstane receptor expression in human hepatocytes: consequences on cytochrome P450 gene regulation. , 2000, Molecular pharmacology.

[35]  K. Yamamoto,et al.  The glucocorticoid receptor inhibits NFkappaB by interfering with serine-2 phosphorylation of the RNA polymerase II carboxy-terminal domain. , 2000, Genes & development.

[36]  J. Pascussi,et al.  Interleukin-6 negatively regulates the expression of pregnane X receptor and constitutively activated receptor in primary human hepatocytes. , 2000, Biochemical and biophysical research communications.

[37]  O. Fardel,et al.  Up‐regulation of multidrug resistance‐associated protein 2 (MRP2) expression in rat hepatocytes by dexamethasone , 1999, FEBS letters.

[38]  D. Häussinger,et al.  Regulation of the multidrug resistance protein 2 in the rat liver by lipopolysaccharide and dexamethasone. , 1999, Gastroenterology.

[39]  P. Thomas,et al.  Induction of the male-specific cytochrome P450 3A2 in female rats by phenytoin. , 1996, Archives of biochemistry and biophysics.

[40]  J. Kountouras,et al.  Prolonged bile duct obstruction: a new experimental model for cirrhosis in the rat. , 1984, British journal of experimental pathology.

[41]  S. Yilmaz,et al.  The possible protective effects of vitamin E and selenium administration in oxidative stress caused by high doses of glucocorticoid administration in the brain of rats. , 2018, Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements.

[42]  A. Xing,et al.  Oxidative stress markers in intrahepatic cholestasis of pregnancy: a prospective controlled study. , 2015, European review for medical and pharmacological sciences.

[43]  B. Staels,et al.  Role of bile acids and bile acid receptors in metabolic regulation. , 2009, Physiological reviews.

[44]  G. Gores,et al.  Toxic bile salts induce rodent hepatocyte apoptosis via direct activation of Fas. , 1999, The Journal of clinical investigation.

[45]  W. Stremmel,et al.  Effect of tauroursodeoxycholic acid on bile-acid-induced apoptosis and cytolysis in rat hepatocytes. , 1998, Journal of hepatology.

[46]  R. Browne,et al.  Reduced glutathione and glutathione disulfide. , 1998, Methods in molecular biology.