Tissue factor-dependent coagulation contributes to α-naphthylisothiocyanate-induced cholestatic liver injury in mice

Abstract

Separation of concentrated bile acids from hepatic parenchymal cells is a key function of the bile duct epithelial cells (BDECs) that form intrahepatic bile ducts. Using coimmunostaining, we found that tissue factor (TF), the principal activator of coagulation, colocalized with cytokeratin 19, a marker of BDECs in the adult mouse liver. BDEC injury induced by xenobiotics such as alpha-naphthylisothiocyanate (ANIT) causes cholestasis, inflammation, and hepatocellular injury. We tested the hypothesis that acute ANIT-induced cholestatic hepatitis is associated with TF-dependent activation of coagulation and determined the role of TF in ANIT hepatotoxicity. Treatment of mice with ANIT (60 mg/kg) caused multifocal hepatic necrosis and significantly increased serum biomarkers of cholestasis and hepatic parenchymal cell injury. ANIT treatment also significantly increased liver TF expression and activity. ANIT-induced activation of the coagulation cascade was shown by increased plasma thrombin-antithrombin levels and significant deposition of fibrin within the necrotic foci. ANIT-induced coagulation and liver injury were reduced in low-TF mice, which express 1% of normal TF levels. The results indicate that ANIT-induced liver injury is accompanied by TF-dependent activation of the coagulation cascade and that TF contributes to the progression of injury during acute cholestatic hepatitis.

Fibrinogen Deficiency Increases Liver Injury and Early Growth Response-1 (Egr-1) Expression in a Model of Chronic Xenobiotic-Induced Cholestasis

“…17,20 However, few studies have determined the exact role of fibrin in cholestatic liver injury. Inhibition of coagulation cascade activation often reduces both hepatic injury and fibrin deposition in models of liver injury, 26,29 -31 leaving many to speculate that fibrin deposition is pathological.…”

“…In the latter two models, substantial periportal necrosis is evident, and these damaged areas are filled with dense fibrin. 16,17,20 It is possible that this fibrin is generated as a consequence of the hepatocellular injury. Indeed, necrosis and fibrin deposition were reduced in livers of PAI-1 Ϫ/Ϫ mice after BDL, although activity of the fibrinolytic enzyme plasmin was unaltered.…”

“…17 For all immunofluorescent staining, 5-m frozen sections obtained from livers frozen in isopentane/OCT were used. For Fbg and cytokeratin 19 (CK19) staining, sections were fixed in 4% neutral-buffered formalin containing 2% acetic acid and subsequently blocked with 10% goat serum in PBS for 1 hour at room temperature.…”

“…[13][14][15] Similarly, coagulation cascade activation occurs in mice subjected to bile duct ligation (BDL), 16 a model of obstructive cholestasis, and in mice give a large dose of ANIT, 17 which models massive intrahepatic bile duct epithelial cell injury. Several studies indicate that plasminogen activator inhibitor-1 (PAI-1) expression increases after BDL and contributes to the progression of obstructive cholestasis in mice.…”

Protective Effects of Moutan Cortex Radicis against Acute Hepatotoxicity

“…Instability of the ANIT-glutathione conjugate and recycling rounds of ANIT metabolism result in high ANIT concentration in bile (Luyendyk et al, 2009). This causes bile duct epithelial cell damage and formation of foci of hepatic necrosis characterized by dead hepatocytes (Luyendyk et al, 2009). …”