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Evaluation of a 3,5-diethoxycarbonyl-1,4-dihydrocollidine diet-induced mouse model in a comparative experimental study of portal hypertension
Jin-Bo Zhao, Zheng-Hao Wu, Jia-Yun Lin, Gu-Qing Luo, Chi-Hao Zhang, Guang-Bo Wu, Qiang Fan, Xiao-Liang Qi, Hai-Zhong Huo, Ji-Wei Yu, Hong-Jie Li, Lei Zheng, Meng Luo
Jin-Bo Zhao, Zheng-Hao Wu, Jia-Yun Lin, Gu-Qing Luo, Chi-Hao Zhang, Guang-Bo Wu, Qiang Fan, Xiao-Liang Qi, Hong-Jie Li, Lei Zheng, Meng Luo, Department of General Surgery, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China
Hai-Zhong Huo, Department of General Surgery, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai 200011, China
Ji-Wei Yu, Department of General Surgery, Ninth People’s Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 201999, China
Co-first authors: Jin-Bo Zhao and Zheng-Hao Wu.
Co-corresponding authors: Lei Zheng and Meng Luo.
Author contributions: Zhao JB and Wu ZH contributed equally to this work; Zhao JB participated in experimental design, conducted core laboratory experiments, and drafted the initial manuscript; Wu ZH handled data collection, statistical analysis, and result validation; Lin JY led literature review and methodology refinement; Luo GQ assisted with experimental reproducibility and manuscript revision; Zhang CH managed sample preparation and quality control; Wu GB and Fan Q collaborated on data visualization and result presentation; Qi XL refined experimental protocols; Huo HZ and Yu JW contributed to manuscript revision and logical flow improvements; Li HJ, Zheng L, and Luo M supervised the project, provided research guidance, secured funding, finalized the manuscript, and approved its submission as corresponding authors; all authors have read and approved the final manuscript.
Supported by the Postdoctoral Scientific Research Foundation of Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, No. 202401023; Shanghai Municipal Commission of Health and Family Planning, No. 20244Y0195 and No. 20234Y0132; and National Natural Science Foundation of China, No. 82100639, No. 82200630 and No. 81970526.
Institutional review board statement: This study does not involve any human experiments.
Institutional animal care and use committee statement: The animal experimental protocol involved in this study has been reviewed and approved by the Ethics Committee of Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University (approval No. SH9H-2021-A233-SB). The experiment was conducted in strict compliance with animal ethics norms and relevant management regulations.
Conflict-of-interest statement: The authors declare that they have 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 original data and related materials can be obtained upon reasonable request made to the corresponding authors.
Corresponding author: Meng Luo, MD, Chief Physician, Department of General Surgery, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Jiao Tong University, No. 639 Manufacturing Bureau Road, Shanghai 200011, China.
luosh9hospital@sina.com
Received: September 16, 2025
Revised: November 6, 2025
Accepted: December 11, 2025
Published online: March 7, 2026
Processing time: 167 Days and 0.1 Hours
BACKGROUND
Portal hypertension (PHT) is a life-threatening complication of chronic liver disease, necessitating reliable animal models that mimic its clinical heterogeneity. Classical mouse models like bile duct ligation (BDL) exhibit a low 4-week survival (35%), while carbon tetrachloride (CCl4) models have delayed pathogenesis, requiring ≥ 8 weeks for PHT development, limiting their efficiency.
AIM
To evaluate the 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet-induced mouse model as a biliary PHT model, comparing it to BDL and CCl4 models.
METHODS
Mice were assigned to DDC diet, BDL, or CCl4 groups. Assessments included portal pressure, histological examination of biliary fibrosis and hepatic stellate cell (HSC) activation (desmin expression), scanning electron microscopy for sinusoidal fenestrae and capillarization, endothelial nitric oxide synthase (eNOS) regulation (phosphorylated-eNOS/total eNOS ratio and total eNOS level), ductular reaction, inflammatory infiltration, and portosystemic shunting. Survival rates and operational feasibility (feed-based administration) were evaluated.
RESULTS
At 4 weeks, DDC induced portal pressure comparable to BDL and CCl4. The DDC model showed moderate biliary fibrosis (similar to BDL but less than CCl4) and greater HSC activation than the other two models. Sinusoidal fenestrae reduction and capillarization in DDC matched BDL and CCl4 models. DDC had decreased phosphorylated-eNOS/total eNOS ratio, while BDL and CCl4 models exhibited reduced total eNOS. DDC demonstrated robust ductular response, inflammation, and shunting, a hallmark of PHT. Survival was 100% (vs 35% BDL, 58.3%-66.6% CCl4), with simpler feed-based induction.
CONCLUSION
The DDC model offers strong biliary PHT relevance, high survival, and efficiency, making it a superior alternative to BDL and CCl4 models for biliary PHT research.
Core Tip: This study validates the 0.1% 3,5-diethoxycarbonyl-1,4-dihydrocollidine diet mouse model for biliary-related portal hypertension (PHT). It matches bile duct ligation (BDL) and carbon tetrachloride (CCl4) models in PHT pathologies (e.g., elevated portal pressure, sinusoidal changes) but outperforms them with 100% 4-week survival (vs 35% BDL, 58.3%-66.6% CCl4), simpler feed-based use, and better recapitulation of PHT hallmarks, serving as a superior alternative.
