Inhibition of nitric oxide synthesis during induced cholestasis ameliorates hepatocellular injury by facilitating S-nitrosothiol homeostasis.
Cholestatic liver injury following extra- or intrahepatic bile duct obstruction causes nonparenchymal cell proliferation and matrix deposition leading to end-stage liver disease and cirrhosis. In cholestatic conditions, nitric oxide (NO) is mainly produced by a hepatocyte-inducible NO synthase (iNOS) as a result of enhanced inflow of endotoxins to the liver and also by accumulation of bile salts in hepatocytes and subsequent hepatocellular injury. This study was aimed to investigate the role of NO and S-nitrosothiol (SNO) homeostasis in the development of hepatocellular injury during cholestasis induced by bile duct ligation (BDL) in rats. Male Wistar rats (200-250 g) were divided into four groups (n=10 each), including sham-operated (SO), bile duct-ligated (BDL), tauroursodeoxycholic acid (TUDCA, 50 mg/kg) and S-methylisothiourea (SMT, 25 mg/kg) treated. After 7 days, BDL rats showed elevated serum levels of gamma-glutamiltranspeptidase, aspartate aminotransferase, alanine aminotransferase, LDH, and bilirubin, bile duct proliferation and fibrosis, compared with the SO group. TUDCA treatment did not significantly alter these parameters, but the iNOS inhibitor SMT ameliorated hepatocellular injury, as shown by lower levels of circulating hepatic enzymes and bilirubin, and a decreased grade of bile duct proliferation and fibrosis. Both TUDCA and SMT treatments reversed Mrp2 canalicular pump expression to control levels. However, only SMT treatment significantly lowered the increased levels of plasma NO and S-nitrosation (S-nitrosylation) of liver proteins in BDL rats. Moreover, BDL resulted in a reduction of the S-nitrosoglutathione reductase (GSNOR/Adh5) enzymatic activity and a downregulation of the GSNOR/Adh5 mRNA expression that was reverted by SMT, but not TUDCA, treatment. A total of 25 liver proteins, including S-adenosyl methionine synthetase, betaine-homocysteine S-methyltransferase, Hsp90 and protein disulfide isomerase, were found to be S-nitrosated in BDL rats. In conclusion, the inhibition of NO production during induced cholestasis ameliorates hepatocellular injury. This effect is in part mediated by the improvement of cell proficiency in maintaining SNO homeostasis.
López-Sánchez
Laura M
LM
Unidad de Investigación del Hospital Universitario Reina Sofía, Instituto Maimónides de Investigación Biomédica, Córdoba 14004, Spain.
Corrales
Fernando J
FJ
Barcos
Montserrat
M
Espejo
Isabel
I
Muñoz-Castañeda
Juan R
JR
Rodríguez-Ariza
Antonio
A
D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors; D000445:Aldehyde Oxidoreductases / Q000037:antagonists & inhibitors / Q000235:genetics; D000818:Animals; D001652:Bile Ducts; D002756:Cholagogues and Choleretics / Q000494:pharmacology; D002779:Cholestasis / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D015536:Down-Regulation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006706:Homeostasis / Q000187:drug effects*; D007550:Isothiuronium / Q000031:analogs & derivatives / Q000494:pharmacology; D008026:Ligation; D008099:Liver / Q000378:metabolism* / Q000473:pathology*; D008297:Male; D009569:Nitric Oxide / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000097:blood; D015538:Nitrosation / Q000187:drug effects; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D026403:S-Nitrosothiols / Q000378:metabolism*; D013655:Taurochenodeoxycholic Acid / Q000494:pharmacology
D018528:ATP-Binding Cassette Transporters; C423013:Abcc2 protein, rat; D002756:Cholagogues and Choleretics; D004791:Enzyme Inhibitors; D011506:Proteins; D012333:RNA, Messenger; D026403:S-Nitrosothiols; D007550:Isothiuronium; D009569:Nitric Oxide; D013655:Taurochenodeoxycholic Acid; C031655:ursodoxicoltaurine; D000445:Aldehyde Oxidoreductases; C087006:formaldehyde dehydrogenase, glutathione-independent; C027744:S-methylisothiopseudouronium