1. Volume 114, Issue 1, Pages 175-184 (January 1998)
Down-regulation of hepatic and renal 11β-hydroxysteroid dehydrogenase in rats with liver cirrhosis 
Geneviève Escher, Andrea Nawrocki, Thomas Staub, Bannikuppe S. Vishwanath, Brigitte M. Frey, Jürg Reichen, Felix J. Frey 
Gastroenterology 
Volume 114, Issue 1, Pages (January 1998)
DOI: /S (98)
Copyright © 1998 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Impact of bile duct ligation on 11β-OHSD1 and 11β-OHSD2 activity in kidney and liver tissues. (A) 11β-OHSD1 oxidase, (B) 11β-OHSD1 reductase, and (C) 11β-OHSD2 oxidase activities were measured and expressed as picomoles or femtomoles of product formed per microgram of total protein, using appropriate substrate concentrations and cofactors. No 11β-OHSD2 activity was found in liver extracts of control or cirrhotic rats, and only 11β-OHSD2 oxidative, but not reductive, activity was found in kidney tissue. ■, Values (±SD) from control animals; ▨, values from cirrhotic animals. 11β-OHSD1 oxidative activity was decreased in kidney (P ≤ 0.043) and liver tissue (P ≤ 0.035) after induction of liver cirrhosis, whereas 11β-OHSD1 reductive activity remained unchanged. 11β-OHSD2 activity was lower (P ≤ 0.01) in cirrhotic kidney tissue extracts. Asterisk indicates statistical differences.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

3. Fig. 1
Impact of bile duct ligation on 11β-OHSD1 and 11β-OHSD2 activity in kidney and liver tissues. (A) 11β-OHSD1 oxidase, (B) 11β-OHSD1 reductase, and (C) 11β-OHSD2 oxidase activities were measured and expressed as picomoles or femtomoles of product formed per microgram of total protein, using appropriate substrate concentrations and cofactors. No 11β-OHSD2 activity was found in liver extracts of control or cirrhotic rats, and only 11β-OHSD2 oxidative, but not reductive, activity was found in kidney tissue. ■, Values (±SD) from control animals; ▨, values from cirrhotic animals. 11β-OHSD1 oxidative activity was decreased in kidney (P ≤ 0.043) and liver tissue (P ≤ 0.035) after induction of liver cirrhosis, whereas 11β-OHSD1 reductive activity remained unchanged. 11β-OHSD2 activity was lower (P ≤ 0.01) in cirrhotic kidney tissue extracts. Asterisk indicates statistical differences.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

4. Fig. 1
Impact of bile duct ligation on 11β-OHSD1 and 11β-OHSD2 activity in kidney and liver tissues. (A) 11β-OHSD1 oxidase, (B) 11β-OHSD1 reductase, and (C) 11β-OHSD2 oxidase activities were measured and expressed as picomoles or femtomoles of product formed per microgram of total protein, using appropriate substrate concentrations and cofactors. No 11β-OHSD2 activity was found in liver extracts of control or cirrhotic rats, and only 11β-OHSD2 oxidative, but not reductive, activity was found in kidney tissue. ■, Values (±SD) from control animals; ▨, values from cirrhotic animals. 11β-OHSD1 oxidative activity was decreased in kidney (P ≤ 0.043) and liver tissue (P ≤ 0.035) after induction of liver cirrhosis, whereas 11β-OHSD1 reductive activity remained unchanged. 11β-OHSD2 activity was lower (P ≤ 0.01) in cirrhotic kidney tissue extracts. Asterisk indicates statistical differences.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

5. Fig. 2
Transcription and translation of 11β-OHSD1 and 11β-OHSD2 in rat liver and kidney tissue. (A) Agarose gel electrophoresis of RT-PCR products of 11β-OHSD1, 11β-OHSD2, and GAPDH in liver and kidney tissue from control and cirrhotic animals. Lane 1, PCR blank (no cDNA); lane 2, liver control; lane 3, liver cirrhotic; lane 4, kidney control; lane 5, kidney cirrhotic; lane 6, molecular weight marker. (B) Western blot of 11β-OHSD1 and 11β-OHSD2 in liver and kidney tissue from control and cirrhotic animals. 11β-OHSD1: lane 1, liver control; lane 2, liver cirrhotic; lane 3, kidney control; lane 4, kidney cirrhotic. 11β-OHSD2: lane 1, kidney control; lane 2, kidney cirrhotic.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

6. Fig. 2
Transcription and translation of 11β-OHSD1 and 11β-OHSD2 in rat liver and kidney tissue. (A) Agarose gel electrophoresis of RT-PCR products of 11β-OHSD1, 11β-OHSD2, and GAPDH in liver and kidney tissue from control and cirrhotic animals. Lane 1, PCR blank (no cDNA); lane 2, liver control; lane 3, liver cirrhotic; lane 4, kidney control; lane 5, kidney cirrhotic; lane 6, molecular weight marker. (B) Western blot of 11β-OHSD1 and 11β-OHSD2 in liver and kidney tissue from control and cirrhotic animals. 11β-OHSD1: lane 1, liver control; lane 2, liver cirrhotic; lane 3, kidney control; lane 4, kidney cirrhotic. 11β-OHSD2: lane 1, kidney control; lane 2, kidney cirrhotic.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

7. Fig. 3
Quantification of steady-state 11β-OHSD1 and 11β-OHSD2 mRNA, using GAPDH as an internal standard. (A) Lower relative amounts of 11β-OHSD1 mRNA were found in control kidney than in control liver tissue (■). The 11β-OHSD1/GAPDH mRNA ratio (A) was significantly lower in cirrhotic liver tissue (▨) than in control tissue (■; P ≤ 0.007), whereas this ratio was not significantly changed by cirrhosis in kidney tissues. A decreased ratio of 11β-OHSD2/GAPDH mRNA (B) was observed in kidney tissues after bile duct ligation (P ≤ 0.046). No signal was found for 11β-OHSD2 mRNA in liver tissue. Asterisk indicates statistical differences.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

8. Fig. 3
Quantification of steady-state 11β-OHSD1 and 11β-OHSD2 mRNA, using GAPDH as an internal standard. (A) Lower relative amounts of 11β-OHSD1 mRNA were found in control kidney than in control liver tissue (■). The 11β-OHSD1/GAPDH mRNA ratio (A) was significantly lower in cirrhotic liver tissue (▨) than in control tissue (■; P ≤ 0.007), whereas this ratio was not significantly changed by cirrhosis in kidney tissues. A decreased ratio of 11β-OHSD2/GAPDH mRNA (B) was observed in kidney tissues after bile duct ligation (P ≤ 0.046). No signal was found for 11β-OHSD2 mRNA in liver tissue. Asterisk indicates statistical differences.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

9. Fig. 4
Inhibition of 11β-OHSD1 oxidative and reductive and 11β-OHSD2 oxidative activity by bile. COS-1 cells were transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 and extracted with Tris/EDTA/Triton or Tris/sucrose buffer for measurement of the corresponding 11β-OHSD activities. Ten micrograms of protein from cell extracts was incubated for 1 hour together with the indicated percentage of bile. Values are means ± SD. (A) Eighty to ninety percent of inhibition was obtained for 11β-OHSD1 (A) oxidative and (B) reductive activity. (C) 11β-OHSD2 activity was enhanced at low concentrations but inhibited at high concentrations of bile.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

10. Fig. 4
Inhibition of 11β-OHSD1 oxidative and reductive and 11β-OHSD2 oxidative activity by bile. COS-1 cells were transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 and extracted with Tris/EDTA/Triton or Tris/sucrose buffer for measurement of the corresponding 11β-OHSD activities. Ten micrograms of protein from cell extracts was incubated for 1 hour together with the indicated percentage of bile. Values are means ± SD. (A) Eighty to ninety percent of inhibition was obtained for 11β-OHSD1 (A) oxidative and (B) reductive activity. (C) 11β-OHSD2 activity was enhanced at low concentrations but inhibited at high concentrations of bile.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

11. Fig. 4
Inhibition of 11β-OHSD1 oxidative and reductive and 11β-OHSD2 oxidative activity by bile. COS-1 cells were transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 and extracted with Tris/EDTA/Triton or Tris/sucrose buffer for measurement of the corresponding 11β-OHSD activities. Ten micrograms of protein from cell extracts was incubated for 1 hour together with the indicated percentage of bile. Values are means ± SD. (A) Eighty to ninety percent of inhibition was obtained for 11β-OHSD1 (A) oxidative and (B) reductive activity. (C) 11β-OHSD2 activity was enhanced at low concentrations but inhibited at high concentrations of bile.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

12. Fig. 5
Screening of various bile salts on the inhibition of 11β-OHSD1 and 11β-OHSD2 activity. COS-1 cells were transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 and were used to assess the inhibitory effect of different bile salts on 11β-OHSD1 (A) oxidative and (B) reductive activity and (C) 11β-OHSD2 activity. Ten micrograms of protein was used for the assay. Each bile salt (50 μmol/L, ■; 500 μmol/L, ▨) was incubated with the reaction mixture. GCA, glycocholic acid; TCDCA, taurochenodeoxycholic acid; GCDCA, glycochenodeoxycholic acid; TDCA, taurodeoxycholic acid; TUDCA, tauroursodeoxycholic acid; GLCA, glycolithocholic acid.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

13. Fig. 5
Screening of various bile salts on the inhibition of 11β-OHSD1 and 11β-OHSD2 activity. COS-1 cells were transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 and were used to assess the inhibitory effect of different bile salts on 11β-OHSD1 (A) oxidative and (B) reductive activity and (C) 11β-OHSD2 activity. Ten micrograms of protein was used for the assay. Each bile salt (50 μmol/L, ■; 500 μmol/L, ▨) was incubated with the reaction mixture. GCA, glycocholic acid; TCDCA, taurochenodeoxycholic acid; GCDCA, glycochenodeoxycholic acid; TDCA, taurodeoxycholic acid; TUDCA, tauroursodeoxycholic acid; GLCA, glycolithocholic acid.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

14. Fig. 5
Screening of various bile salts on the inhibition of 11β-OHSD1 and 11β-OHSD2 activity. COS-1 cells were transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 and were used to assess the inhibitory effect of different bile salts on 11β-OHSD1 (A) oxidative and (B) reductive activity and (C) 11β-OHSD2 activity. Ten micrograms of protein was used for the assay. Each bile salt (50 μmol/L, ■; 500 μmol/L, ▨) was incubated with the reaction mixture. GCA, glycocholic acid; TCDCA, taurochenodeoxycholic acid; GCDCA, glycochenodeoxycholic acid; TDCA, taurodeoxycholic acid; TUDCA, tauroursodeoxycholic acid; GLCA, glycolithocholic acid.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

15. Fig. 6
Concentration-response curves of TCA and CDCA vs. percentage of inhibition of 11β-OHSD1 and 11β-OHSD2 from extracts of transfected COS-1 cells. Cell extracts of COS-1 transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 were incubated with TCA or CDCA, respectively. Values are means ± SD. TCA inhibited 11β-OHSD1 oxidative activity (A) with an inhibition constant (Ki) of 6.7 μmol/L and inhibited reductive activity (B) with a Ki of 7.7 μmol/L. CDCA inhibited 11β-OHSD2 activity (C) with a Ki of 22.7 μmol/L.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

16. Fig. 6
Concentration-response curves of TCA and CDCA vs. percentage of inhibition of 11β-OHSD1 and 11β-OHSD2 from extracts of transfected COS-1 cells. Cell extracts of COS-1 transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 were incubated with TCA or CDCA, respectively. Values are means ± SD. TCA inhibited 11β-OHSD1 oxidative activity (A) with an inhibition constant (Ki) of 6.7 μmol/L and inhibited reductive activity (B) with a Ki of 7.7 μmol/L. CDCA inhibited 11β-OHSD2 activity (C) with a Ki of 22.7 μmol/L.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

17. Fig. 6
Concentration-response curves of TCA and CDCA vs. percentage of inhibition of 11β-OHSD1 and 11β-OHSD2 from extracts of transfected COS-1 cells. Cell extracts of COS-1 transfected with the cDNA of 11β-OHSD1 or 11β-OHSD2 were incubated with TCA or CDCA, respectively. Values are means ± SD. TCA inhibited 11β-OHSD1 oxidative activity (A) with an inhibition constant (Ki) of 6.7 μmol/L and inhibited reductive activity (B) with a Ki of 7.7 μmol/L. CDCA inhibited 11β-OHSD2 activity (C) with a Ki of 22.7 μmol/L.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions