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Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis

  • Authors:
    • Wanrong Xia
    • Hongji Yao
    • Tongxin Mu
    • Hui Liu
    • Jianhua Chen
    • Shi Feng
    • Shilei Wen
  • View Affiliations / Copyright

    Affiliations: Key Laboratory of Tropical Translational Medicine of Ministry of Education and Key Laboratory of Brain Science Research Transformation in Tropical Environment of Hainan Province, School of Basic Medicine and Life Sciences, Hainan Medical University, Haikou, Hainan 571199, P.R. China, Department of Forensic Medicine, College of Basic Medicine and Life Sciences, Hainan Medical University, Haikou, Hainan 571199, P.R. China, Department of Histology and Embryology, Basic Medical School, Kunming University of Science and Technology, Kunming, Yunnan 650500, P.R. China
    Copyright: © Xia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 167
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    Published online on: July 3, 2025
       https://doi.org/10.3892/etm.2025.12917
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Abstract

Common bile duct ligation (BDL) in rats is a widely used animal model for the study of cholestatic liver injury; however, it has a high mortality rate and leads to rapid disease progression. To address these limitations, the present study developed a simplified partial BDL (p‑BDL) procedure by inserting a 26G disposable venous indwelling needle during ligation of the common bile duct, and subsequently removing the needle to create a narrow passage through the ligature points. The rats were euthanized 1 or 2 months after the procedure for histological, immunohistochemical and gelatin zymography analyses of the liver. Notably, no animals died during the experimental period, demonstrating the low mortality rate of the model. When assessed 1 month post‑p‑BDL, the rats exhibited mild liver fibrosis, while at 2 months, severe liver fibrosis was observed. Corresponding changes in liver function indices were also noted. Specifically, markers of liver fibrosis, including collagen I, collagen III and α‑smooth muscle actin, as well as matrix metalloproteinase (MMP)‑2 and MMP‑9, which contribute to extracellular matrix degradation, were significantly upregulated following p‑BDL. These findings are similar to those observed following BDL, and indicate that the p‑BDL model was successfully established. This model mitigates the drawbacks of traditional BDL and mimics chronic cholestatic cirrhosis, as observed clinically.
View Figures

Figure 1

Representative images of the partial
common bile duct ligation procedure. (A) Rat bound in a supine
position; (B) skin preparation and disinfection; (C) opening of the
abdominal cavity; (D) exposure of the bile duct; (E) insertion of
the surgical sutures; (F) binding the trocar; (G) pulling out the
needle core and cutting off the cannula; (H) pushing the intestine
back into the abdomen; (I) closing the abdomen layer by layer.

Figure 2

Gross appearance and liver function
indices of the rats following partial common bile duct ligation.
(A) Photographs of a representative rat from each group. (B)
Adhesion between the inferior margin of the liver and the bowel.
(C) Enlargement of the common bile duct. Serum levels of (D) ALT,
(E) AST and (F) TBIL. All values are presented as the mean ±
standard deviation (n=10). **P<0.01 vs. sham group;
##P<0.01 vs. 1 MPO group. ALT, alanine
aminotransferase; AST, aspartate aminotransferase; TBIL, total
bilirubin; MPO, month post operation.

Figure 3

Histopathological changes of rat
livers following partial common bile duct ligation. Representative
images and histopathological analysis by (A) H&E staining, (B)
Sirius red staining and (C) Masson's trichrome staining (scale bar,
200 µm; magnification, x100). (D) CK-19 immunostaining (scale bar,
100 µm; magnification, x100). (E) Ishak's scores of hepatic
fibrosis based on the staining results. Blue dashed lines demarcate
fibrotic tissue boundary. Red arrows demarcate pseudo bile ducts in
Fig. 3D. All values are presented
as the mean ± standard deviation (n=10). *P<0.05,
**P<0.01 or ***P<0.001 vs. the sham
group; #P<0.05 or ##P<0.01 vs. the 1
MPO group. H&E, hematoxylin and eosin; CK-19, cytokeratin 19;
MPO, month post operation.

Figure 4

Upregulation of fibrotic markers and
MMPs in rat livers following partial common bile duct ligation.
Representative IHC staining images and quantification of (A)
collagen I, (B) collagen III and (C) α-SMA. Representative IHC
staining images of (D) MMP-2 and (E) MMP-9. (A-E) Scale bar, 100
µm; magnification, x100. (F) Representative gelatin zymography
results and (G) the relative activities of MMP complex, pro-MMP-2,
active MMP-2 and MMP-9 in the rat livers. All values are presented
as mean ± standard deviation (n=5). *P<0.05 or
**P<0.01 vs. the sham group; ##P<0.01
vs. the 1 MPO group. MMP, matrix metalloproteinase; IHC,
immunochemistry; SMA, smooth muscle actin; MPO, month post
operation.
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Copy and paste a formatted citation
Spandidos Publications style
Xia W, Yao H, Mu T, Liu H, Chen J, Feng S and Wen S: Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis. Exp Ther Med 30: 167, 2025.
APA
Xia, W., Yao, H., Mu, T., Liu, H., Chen, J., Feng, S., & Wen, S. (2025). Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis. Experimental and Therapeutic Medicine, 30, 167. https://doi.org/10.3892/etm.2025.12917
MLA
Xia, W., Yao, H., Mu, T., Liu, H., Chen, J., Feng, S., Wen, S."Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis". Experimental and Therapeutic Medicine 30.3 (2025): 167.
Chicago
Xia, W., Yao, H., Mu, T., Liu, H., Chen, J., Feng, S., Wen, S."Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis". Experimental and Therapeutic Medicine 30, no. 3 (2025): 167. https://doi.org/10.3892/etm.2025.12917
Copy and paste a formatted citation
x
Spandidos Publications style
Xia W, Yao H, Mu T, Liu H, Chen J, Feng S and Wen S: Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis. Exp Ther Med 30: 167, 2025.
APA
Xia, W., Yao, H., Mu, T., Liu, H., Chen, J., Feng, S., & Wen, S. (2025). Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis. Experimental and Therapeutic Medicine, 30, 167. https://doi.org/10.3892/etm.2025.12917
MLA
Xia, W., Yao, H., Mu, T., Liu, H., Chen, J., Feng, S., Wen, S."Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis". Experimental and Therapeutic Medicine 30.3 (2025): 167.
Chicago
Xia, W., Yao, H., Mu, T., Liu, H., Chen, J., Feng, S., Wen, S."Novel partial common bile duct ligation procedure in rats with reduced mortality and delayed onset of cirrhosis". Experimental and Therapeutic Medicine 30, no. 3 (2025): 167. https://doi.org/10.3892/etm.2025.12917
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