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Key genes and histopathological alterations in hepatic fibrogenesis after segmental cholestasis in weaned rats

Pediatric Research (2025)Cite this article

Abstract

Background

The selective bile duct ligation (sBDL) model has been proposed to study fibrosis progression in segmental cholestasis, replicating histopathological features of biliary obstruction in both liver parenchyma with biliary obstruction (BO) and without biliary obstruction (WBO). However, the molecular mechanisms driving fibrogenesis in WBO parenchyma remain unclear. This study aimed to characterize gene expression and histological alterations in fibrogenesis using the sBDL model.

Methods

Liver samples were collected at 1, 4, and 8 weeks post-surgery from both BO and WBO parenchyma, undergoing histological, immunohistochemical, biochemical, and molecular analyses.

Results

Differentially expressed genes (DEGs) were associated to inflammatory response, extracellular matrix production, angiogenesis, and negative regulation of peptidase activity. Histologically, ductular proliferation, inflammatory infiltration, and collagen deposition were observed in both BO and WBO parenchyma, with more pronounced inflammation and hepatocellular degeneration in BO. BO parenchyma showed more pathways related to disease progression. In contrast, pathways related to cellular senescence and the PI3K-Akt signaling, suggesting suppression of apoptosis and cell proliferation.

Conclusion

The sBDL model mirrors key aspects of human biliary fibrosis, offering novel molecular insights into fibrogenesis in segmental cholestasis and serving as a valuable tool for developing diagnostic and therapeutic strategies.

Impact

  • This study demonstrates that hepatic fibrogenesis extends beyond regions directly affected by biliary obstruction, involving non-obstructed liver parenchyma. Using the selective bile duct ligation (sBDL) model, we identified key molecular pathways associated with fibrogenesis in segmental cholestasis. Our findings provides new insights into the mechanisms of cholestatic fibrosis and highlighting the sBDL model as a valuable preclinical tool. These results may inform the development of novel therapeutic strategies for treating biliary fibrosis.

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Fig. 1: Post-surgical clinical evaluation.
Fig. 2: Representative photomicrographs of histological and immunohistochemical analyses in sham, BO, and WBO groups.
Fig. 3: Graphical representation of the analyses of histological and biochemical assessments of liver injury and fibrosis.
Fig. 4: Representative photomicrographs of Ki-67 immunohistochemistry for cell proliferation quantification (brown) and cleaved caspase-3 immunohistochemistry for apoptosis analysis (brown).
Fig. 5: Gene Expression Analysis.
Fig. 6: Validation of RT-PCR array experiments by quantitative real-time PCR (qRT-PCR) for selected genes.
Fig. 7: Pearson correlation analysis of genes, GO BP terms, and KEGG pathways with phenotypic changes observed in histopathological features.

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Data availability

All data generated or analyzed during this study are included in this article. The datasets used during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This study was funded by São Paulo Research Foundation—FAPESP, (Project Number: 2015/00977-0).

Author information

Authors and Affiliations

  1. Division of Pediatric Surgery, Pediatric Liver Transplant Unit, and Pediatric Surgery Research Laboratory (LIM 30), University of São Paulo School of Medicine, São Paulo, SP, Brazil

    Josiane O. Gonçalves, Suellen Serafini, Uenis Tannuri & Ana C. A. Tannuri

  2. Department of Pediatrics, University of São Paulo School of Medicine, São Paulo, SP, Brazil

    Josiane O. Gonçalves, Silvia Y. Bando, Suellen Serafini, Carlos A. Moreira-Filho, Uenis Tannuri & Ana C. A. Tannuri

  3. Liver and Gastrointestinal Transplant Division, Department of Gastroenterology, University of São Paulo School of Medicine, São Paulo, SP, Brazil

    Cleiton A. M. Mafra

  4. Department of Nephrology, University of São Paulo School of Medicine, São Paulo, SP, Brazil

    Isac de Castro

  5. Department of Pathology, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, SP, Brazil

    Bruno Cogliati

  6. Division of Rheumatology, Hospital das Clínicas, University of São Paulo School of Medicine, São Paulo, SP, Brazil

    Walcy R. Teodoro

Authors
  1. Josiane O. Gonçalves
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  10. Ana C. A. Tannuri
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Contributions

J.O. Gonçalves and A.C.A. Tannuri designed the study. J.O. Gonçalves, C.A.M. Mafra, W.R. Teodoro, Cogliati B, and Serafini S conducted the experiments and collected the data; B Cogliati performed the histopathological evaluation; J.O. Gonçalves, I. de Castro, S.Y. Bando, and C.A. Moreira-Filho analyzed and interpreted the data; J.O. Gonçalves, C.A.M. Mafra, and A.C.A. Tannuri wrote the manuscript; all authors approved the final draft of the manuscript; the acquisition of funding was completed by U. Tannuri and A.C.A. Tannuri.

Corresponding author

Correspondence to Josiane O. Gonçalves.

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Gonçalves, J.O., Mafra, C.A.M., Castro, I.d. et al. Key genes and histopathological alterations in hepatic fibrogenesis after segmental cholestasis in weaned rats. Pediatr Res (2025). https://doi.org/10.1038/s41390-025-04272-x

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