Adjunctive Cystic Duct Ligation Refines the Murine Bile Duct Ligation Model of Portal Hypertension

Background Murine bile duct ligation (BDL) is widely used to model cholestatic liver injury and portal hypertension (PH), but postoperative complications and mortality undermine its experimental utility. We examined whether adjunctive cystic duct ligation (CDL) improves survival while preserving the portal hypertensive phenotype. Male C57BL/6J mice underwent sham surgery, conventional BDL, or the refined BDL + CDL procedure. Four-week survival was compared among groups. Portal pressure was assessed in independent terminal cohorts at 1, 2, 3, and 4 weeks post-surgery. Systemic hemodynamics, organ-to-body weight ratios, liver fibrosis, and biochemical injury markers were comprehensively evaluated. Results The refined BDL + CDL group showed markedly improved postoperative survival at four weeks compared with the BDL group (56.1% vs. 28.3%; hazard ratio, 0.490; 95% confidence interval, 0.304–0.790; log-rank P = 0.003). BDL + CDL reduced the incidence of postoperative complications, particularly gallbladder perforation and ascites. In addition, the BDL + CDL procedure induced a more severe and progressive PH, with portal pressure significantly higher than that in the BDL group. The exacerbated PH was associated with aggravated intrahepatic fibrosis, evidenced by advanced histopathological staging, increased collagen deposition, pro-fibrogenic molecular signature, and elevated liver injury markers. Mean arterial pressure and heart rate did not differ significantly among groups. Conclusions Incorporating CDL into the standard BDL procedure establishes a more robust and reproducible experimental model for studying PH. This refined model offers translational utility for investigating PH pathophysiology and evaluating novel anti-fibrotic therapies.