Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Portal pressure responses and angiotensin peptide production in rat liver are determined by relative activity of ACE and ACE2

doi: 10.1152/ajpgi.00045.2009

Figure Lengend Snippet: Angiotensin-(1-7) production in rat liver. Ang I (99 pmol) or Ang II (60 pmol) bolus was injected into the portal vein of in situ perfused healthy (○ and open bars) and fibrotic (• and solid bars) rat liver preparations with or without the ACE inhibitor lisinopril (10−6 mol/l), ACE and NEP inhibitor omapatrilat (10−6 mol/l), ACE2 inhibitor MLN4760 (10−6 mol/l), or a combination of MLN4760 and omapatrilat. Angiotensin-(1-7) production from Ang II (A) and the baseline-corrected total area under the respective curves (B), and angiotensin-(1-7) production from Ang I (C) and the baseline-corrected total area under the respective curves (D), and Ang II-induced portal pressure responses (E) and the area under pressure response curve (F) are shown. Each symbol represents the mean ± SE profile from 4–5 rats per treatment group. Note that y-axis scale is different between A and C and between B and D. Arrow indicates the time of bolus injection. ****P < 0.0005, **P < 0.01, *P < 0.05, vs. respective healthy livers. ††††P < 0.0005, between the 2 groups indicated by brackets. In E and F, *P < 0.05 vs. other groups except bile duct ligation (BDL)/ACE inhibition group (P = 0.059).

Article Snippet: MLN4760 was provided by Millennium Pharmaceuticals, Cambridge, MA.

Techniques: Injection, In Situ, Ligation, Inhibition

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Portal pressure responses and angiotensin peptide production in rat liver are determined by relative activity of ACE and ACE2

doi: 10.1152/ajpgi.00045.2009

Figure Lengend Snippet: Ang II production and portal pressure changes in response to Ang I in rat liver. Ang I bolus (99 pmol) was injected into the portal vein of in situ perfused healthy (○ and open bars) and fibrotic (• and solid bars) rat liver preparations with or without the ACE inhibitor lisinopril (10−6 mol/l), ACE and NEP inhibitor omapatrilat (10−6 mol/l), ACE2 inhibitor MLN4760 (10−6 mol/l), or a combination of MLN4760 and omapatrilat. Ang II production (A) and the area under the Ang II curve (B) and Ang I-induced portal pressure responses (C) and the area under pressure-response curve (D) are shown. Each symbol represents the mean ± SE profile from 4–5 rats per treatment group. Arrows indicate the time of bolus injection. ****P < 0.0005, **P < 0.01, *P < 0.05, vs. respective healthy livers. ††††P < 0.0005, †P < 0.05, between the 2 groups indicated by brackets.

Article Snippet: MLN4760 was provided by Millennium Pharmaceuticals, Cambridge, MA.

Techniques: Injection, In Situ

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Portal pressure responses and angiotensin peptide production in rat liver are determined by relative activity of ACE and ACE2

doi: 10.1152/ajpgi.00045.2009

Figure Lengend Snippet: ACE2 activity of the perfused rat liver. In situ perfused healthy (open bars) and fibrotic (solid bars) rat livers, incubated with or without the ACE inhibitor lisinopril (10−6 mol/l), ACE and NEP inhibitor omapatrilat (10−6 mol/l), ACE2 inhibitor MLN4760 (10−6 mol/l), or a combination of MLN4760 and omapatrilat were used for cell membrane preparations for measurement of ACE2 activity. ACE2 activity was determined by measuring nanomoles of ACE2 substrate [Mca-APK-(Dnp)-OH] cleaved by solubilized membrane fractions. Each bar represents the mean ± SE activity from 4–5 rats per treatment group. ACE2 activity was not different between different groups of healthy livers so that the mean value from all groups is shown. ****P < 0.0005, ***P < 0.005, **P < 0.01, *P < 0.05 vs. group indicated by brackets.

Article Snippet: MLN4760 was provided by Millennium Pharmaceuticals, Cambridge, MA.

Techniques: Activity Assay, In Situ, Incubation, Membrane

Journal: Hypertension

Article Title: Measurement of Angiotensin Converting Enzyme 2 Activity in Biological Fluid (ACE2)

doi: 10.1007/978-1-4939-6625-7_8

Figure Lengend Snippet: Standard curves for ACE2 activity assay. The assay was performed for 16 h incubation. Graph shows the concentrations of recombinant ACE2 (rACE2) versus the ACE2 activity (RFU). ( a ) Standard curves generated for human rACE2. The ACE2 inhibitors MLN-4760 (10 −6 M) or DX600 (10 −6 M) were used. The relationship is highly linear for rACE2 concentrations between 0 and 50 ng/ml ( R 2 = 0.9998, p < 0.001 for MLN-4760, R 2 = 0.9999, p < 0.001 for DX600). ( b ) Standard curve generated for mouse rACE2. The ACE2 inhibitor MLN-4760 (10 −6 M) was used. A highly linear relationship exists for rACE2 concentrations between 0 and 50 ng/ml ( R 2 = 0.9976, p < 0.001)

Article Snippet: 2 inhibitor: MLN-4760 (10 −5 M stock in ddH 2 O, also called GL1001, provided by Ore Pharmaceuticals, Cambridge, MA, USA, and commercially available from Calbiochem, San Diego, CA, USA, Catalog Number: 530616) or DX600 (linear form, 10 −5 M stock in ddH 2 O, AnaSpec, Catalog Number: 62337) .

Techniques: Activity Assay, Incubation, Recombinant, Generated

Journal: Hypertension

Article Title: Measurement of Angiotensin Converting Enzyme 2 Activity in Biological Fluid (ACE2)

doi: 10.1007/978-1-4939-6625-7_8

Figure Lengend Snippet: Effect of ACE2 inhibitors MLN-4760 (10 −6 M), DX600 (10 −6 M), and ACE inhibitor captopril (10 −5 M) on ACE2 enzyme activity. The assay was performed for 16 h incubation. Graph depicts the concentrations of recombinant ACE2 versus the Relative Fluorescence Units (RFU). A background RFU value (67.5 RFU for a , 62.0 RFU for b ) of the substrate blank control was subtracted from the readings for each well. ( a ) Effect of MLN-4760, DX600, and captopril on human rACE2 activity. ( b ) Effect of MLN-4760, DX600, and captopril on mouse rACE2 activity

Article Snippet: 2 inhibitor: MLN-4760 (10 −5 M stock in ddH 2 O, also called GL1001, provided by Ore Pharmaceuticals, Cambridge, MA, USA, and commercially available from Calbiochem, San Diego, CA, USA, Catalog Number: 530616) or DX600 (linear form, 10 −5 M stock in ddH 2 O, AnaSpec, Catalog Number: 62337) .

Techniques: Activity Assay, Incubation, Recombinant, Fluorescence