Lithocholic Acid Feeding Induces Segmental Bile Duct Obstruction and Destructive Cholangitis in Mice

Abstract

We determined the mechanisms of hepatobiliary injury in the lithocholic acid (LCA)-fed mouse, an increasingly used model of cholestatic liver injury. Swiss albino mice received control diet or 1% (w/w) LCA diet (for 1, 2, and 4 days), followed by assessment of liver morphology and ultrastructure, tight junctions, markers of fibrosis and key proteins of hepatobiliary function, and bile flow and composition. As expected LCA feeding led to bile infarcts, which were followed by a destructive cholangitis with activation and proliferation of periductal myofibroblasts. At the ultrastructural level, small bile ducts were frequently obstructed by crystals. Biliary-excreted fluorescence-labeled ursodeoxycholic acid accumulated in bile infarcts, whereas most infarcts did not stain with India ink injected into the common bile duct; both findings are indicative of partial biliary obstruction. Expression of the main basolateral bile acid uptake proteins (sodium-taurocholate cotransporter and organic anion-transporting polypeptide 1) was reduced, the canalicular transporters bile salt export pump and multidrug-related protein 2 were preserved, and the basolateral transporter multidrug-related protein 3 and the detoxifying enzyme sulfotransferase 2a1 were induced. Thus, we demonstrate that LCA feeding in mice leads to segmental bile duct obstruction, destructive cholangitis, periductal fibrosis, and an adaptive transporter and metabolic enzyme response.

A New Xenobiotic-Induced Mouse Model of Sclerosing Cholangitis and Biliary Fibrosis

“…protein and analysis for determination of vascular cell adhesion molecule (VCAM) protein was performed as described. 12 …”

“…Immunohistochemistry for CD11b, proliferation marker Ki-67, and keratin 19 (K19) were performed as described. 12 mRNA analysis and polymerase chain reaction of several genes was performed as described. 12 Primers used for mRNA analysis of key genes for transporters, metabolic enzymes, inflammation, and fibrosis are summarized in Table 2.…”

Constitutive Androstane Receptor-Mediated Changes in Bile Acid Composition Contributes to Hepatoprotection from Lithocholic Acid-Induced Liver Injury in Mice

“…Excess concentrations of bile acids also cause adaptive changes in the liver, such as decreased hepatobiliary transport (Zollner et al, 2003). For example, Fickert et al (2006) have shown that administration of LCA for 4 days in mice can result in hepatocellular necrosis with significant reductions in basolateral bile acid uptake (Ntcp, Oatp1) and increased expression of sinusoidal bile acid efflux transporters (Mrp3). These adaptive changes in the liver represent an attempt to protect cells from the inherent toxicity of accumulating bile acids.…”

3-Ketocholanoic Acid Is the Major in Vitro Human Hepatic Microsomal Metabolite of Lithocholic Acid

“…Treatment of hepatocytes in vitro with lithocholic acid at a concentration of 1 mM increased mitochondrial membrane permeability and caused cellular lysis (Rolo et al, 2000;Palmeira and Rolo, 2004). Administration of lithocholic acid to experimental animals produced liver and biliary tract injury, including bile duct proliferation, multifocal necrosis with vacuolization, inflammation of the portal area, and atrophy of hepatic lobules (Fickert et al, 2006;Beilke et al, 2008). These pathophysiological changes are indicative of intrahepatic cholestasis and cirrhosis (Palmer and Ruban, 1966;Miyai et al, 1971;Fischer et al, 1974).…”