1. Volume 123, Issue 5, Pages 1667-1676 (November 2002)
Up-regulation of components of the renin-angiotensin system in the bile duct–ligated rat liver 
Georgina Paizis, Mark E. Cooper, Josefa M. Schembri, Christos Tikellis, Louise M. Burrell, Peter W. Angus 
Gastroenterology 
Volume 123, Issue 5, Pages (November 2002)
DOI: /gast
Copyright © 2002 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Hematoxylin and eosin preparations of representative liver samples of (A) sham-operated animals showing normal liver histology and (B) bile duct-ligated animals with bile ductular proliferation, expansion of portal tracts, portal to portal fibrous bridging, and nodular transformation.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Quantitation of mRNA by quantitative real-time RT-PCR. Renin (A) and angiotensinogen (B) gene expression was unchanged following injury (P = 0.31 and 0.12, respectively). ACE (C) and AT1 receptor (D) expression were significantly higher in bile duct animals compared with sham-operated animals (*P < 0.005; †P < 0.04, respectively). Data are shown as mean ± SEM for each specific gene relative to the level in sham animals, which has been arbitrarily assigned a value of 1.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Representative autoradiographic mapping of ACE activity in liver sections with red representing high-density binding sites and blue/green low-density binding or background (see scale). Minimal binding is seen in the liver of sham-operated animals (A). ACE binding was increased following bile duct ligation (B). The graph below demonstrates quantitation of ACE activity by densitometry scanning of autoradiographs in the groups of animals represented in the liver sections shown above. Sham livers display 4.5% binding, which is increased to 48.7% following bile duct ligation (*P < 0.001; BDL vs. sham). Data are shown as mean ± SEM.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Localization of ACE in liver sections, with silver grains representing sites of ACE binding or activity. ACE binding is homogeneously distributed in normal liver (A and B, light field and dark field, respectively). Following bile duct ligation, ACE binding is also seen in fibrotic bands. (Original magnification in A, B 200×; in C, D 100×.)
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Localization of the AT1 receptor subtype by ligand-binding studies. Silver grains represent sites of AT1 receptor binding. Heterogeneous signal is seen in hepatocytes of sham liver (A). Following bile duct ligation (B), there is intense signal in specific cells within areas of active fibrogenesis. These eosinophilic cells stain with toluidine blue (insert) and were identified as mast cells. Bile duct cells (BDC) and hepatocytes (H) are identified. Immunostaining of liver sections with α-smooth muscle actin antibody (C) demonstrating the distribution of spindle-shaped hepatic stellate cells within an area of active fibrogenesis and bile duct proliferation (BD). Macrophages are identified with ED1 immunohistochemistry (D), which are present in the sinusoids of liver parenchyma and not in the areas of fibrosis and bile duct proliferation (BD).
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Localization of angiotensinogen (A and B) and ACE (C and D) protein by immunohistochemistry. Angiotensinogen staining is seen in hepatocytes surrounding the central vein (CV) of sham liver (A). Following bile duct ligation (B), angiotensinogen protein is detected in the region of active fibrogenesis, indicating de novo synthesis of this protein in this region. ACE protein is not immunostainable in normal liver (C) but is readily detected in bile duct cells of proliferating bile ducts (BD) following bile duct ligation (D). There is no staining in adjacent hepatocytes (H).
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

8. Fig. 7
Western analysis of renin expression in kidney tissue (positive controls), sham, and BDL livers. Protein extracts of kidney, sham, and BDL liver tissue were probed with a renin-specific antibody. Renin protein is abundant in kidney but expressed in low levels in both sham and BDL livers. The autoradiographs were exposed for 7 minutes.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions