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World J Gastroenterol. Dec 28, 2026; 32(48): 122778
Published online Dec 28, 2026. doi: 10.3748/wjg.122778
Establishment of a Sprague-Dawley rat model of hepatolithiasis based on common bile duct catheterization
Jia-Lin Shi, Tauqeer Muhammad, Ting-Ting He, Zhu-Ying Lin, Tian-Gui Zhang, Shi-Yue Liu, Xiao-Lei Yang, Yao Yang, Lei Hu, Pan Ran, Muhammad Numan, Jiang Li
Jia-Lin Shi, Tauqeer Muhammad, Ting-Ting He, Zhu-Ying Lin, Tian-Gui Zhang, Xiao-Lei Yang, Yao Yang, Lei Hu, Pan Ran, Jiang Li, Department of Hepatobiliary Surgery, The First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, China
Shi-Yue Liu, Department of Pathology, The First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, China
Muhammad Numan, School of Pharmaceutical Sciences & Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming 650500, Yunnan Province, China
Co-first authors: Jia-Lin Shi and Tauqeer Muhammad.
Author contributions: Shi JL conceived and designed the study, conducted the primary research, collected and analyzed the data, drafted the original manuscript, and made critical revisions to the intellectual content; Muhammad T conducted the research and contributed to data collection and analysis; He TT and Lin ZY conducted the literature search and contributed to background documentation and reference compilation; Zhang TG, Liu SY, and Yang XL assisted in data interpretation and contributed to the methodology and validation processes; Yang Y and Hu L made critical revisions to the manuscript and contributed to its intellectual content; Ran P assisted in manuscript preparation and provided essential technical support throughout the study; Numan M contributed to manuscript revision and language polishing; Li J contributed to the conceptualization and design of the study, provided expert guidance throughout all phases of the research, critically reviewed the manuscript, and provided final approval of the version to be published; and all authors reviewed and approved the final submitted version of the manuscript. Shi JL and Muhammad T contributed equally to this study and these collective contributions underlie their merit as co-first authors.
AI contribution statement: AI tools were not used to generate original scientific data, perform independent scientific analyses, or draw scientific conclusions. The authors take full responsibility and accountability for all content of this manuscript, including any portions for which AI tools were not used.
Supported by “Xingdian Talents” Support Project of Yunnan Province.
Institutional review board statement: The study was reviewed and approved by the Ethics Committee of the First Affiliated Hospital of Kunming Medical University (Approval No. 2025 L309).
Institutional animal care and use committee statement: The study was reviewed and approved by the Animal Experimental Ethics Committee of the First Affiliated Hospital of Kunming Medical University (Approval No. 2025DF039).
Conflict-of-interest statement: All authors declared no conflict of interest.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: All data are available on request.
Corresponding author: Jiang Li, PhD, Chief Physician, Full Professor, Department of Hepa
tobiliary Surgery, The First Affiliated Hospital of Kunming Medical University, No. 295 Xichang Road, Wuhua District, Kunming 650032, Yunnan Province, China.
lijiang1020@kmmu.edu.cn
Received: April 28, 2026
Revised: July 22, 2026
Accepted: September 11, 2026
Published online: December 28, 2026
Processing time: 189 Days and 5.5 Hours
BACKGROUND
Hepatolithiasis is a common benign bile duct disease prevalent in East Asian regions, characterized primarily by chronic inflammation of the intrahepatic bile ducts, bile stasis, and the formation of calcium bilirubinate stones. The underlying mechanisms of its development have not yet been fully elucidated. Establishing animal models that can simulate the process of chronic cholangitis and the formation of calcium bilirubinate stones is crucial for elucidating the mechanisms of disease development and evaluating potential interventions.
AIM
To establish and optimize a Sprague-Dawley rat model of hepatolithiasis based on common bile duct catheterization combined with staged lipopolysaccharide injection; evaluate its stability, reproducibility, and histopathological characteristics; and provide a reliable animal model for studying the pathogenesis of hepatolithiasis and evaluating related intervention strategies.
METHODS
Twelve specific pathogen-free-grade male Sprague-Dawley rats weighing 400 ± 25 g were randomly divided into an experimental group and a control group, with 6 rats in each group. After establishing a common bile duct catheterization model, the experimental group received lipopolysaccharide (1 mg/kg) injections into the bile duct via the catheter on postoperative days 4, 9, 14, and 19, while the control group received an equivalent volume of saline at the same time points. The catheter was ligated for 6 hours after each injection. On postoperative day 24, the animals were euthanized, and bile and liver tissue were collected. The composition of bile stones was analyzed through gross observation of bile, microscopic examination of bile smears, and Fourier transform infrared spectroscopy. The degree of cholangitis and fibrosis was evaluated using hematoxylin and eosin staining and Scheuer scoring to comprehensively validate the model.
RESULTS
The bile of the experimental group was darker in color and had greater viscosity. Microscopic examination of bile smears showed crystals that resembled human biliary stones. Fourier transform infrared spectroscopy analysis showed calcium bilirubinate to be the major constituent. Hematoxylin and eosin staining showed significant inflammatory infiltration of the portal tracts, and inflammation scores were higher (G1-G2) than those in the control group (G0-G1). The portal tracts in the experimental group showed fibrotic changes (S1), whereas those in the control group did not (S0).
CONCLUSION
This model recapitulates biliary changes, crystal formation, and the process of cholangitis-fibrosis in multiple dimensions. It offers the practicality of long-term drug delivery and periodic sampling through an indwelling catheter, providing a stable experimental model for examining abnormal bilirubin metabolism, biliary inflammatory responses, and the pathogenesis of stone formation.
Core Tip: This study successfully established a hepatolithiasis model in Sprague-Dawley rats based on bile duct catheterization and intrabiliary administration of lipopolysaccharide (1 mg/kg) on postoperative days 4, 9, 14, and 19. The model was validated using multiple methods, including microscopic examination of bile smears, identification of calcium bilirubinate crystals by Fourier transform infrared spectroscopy, and histopathological assessment using Scheuer scoring. These findings confirmed that the model consistently produces calcium bilirubinate stones and reproduces typical pathological changes such as chronic biliary inflammation and biliary fibrosis. Additionally, this model allows for repeated intrabiliary dosing and continuous bile sampling, making it a reliable experimental platform for studying the pathogenesis of intrahepatic bile duct stones and evaluating novel intervention strategies.
