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5 protocols using ave0991

1

Osmotic mini-pump delivery of AVE0991 in BDL rats

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Osmotic mini-pumps (Model 2002, Alzet, Cupertino, CA, USA) were filled with AVE0991 (a gift from Sanofi-Aventis Deutschland GmbH, Germany) dissolved in polyethylene glycol 400 (Sigma-Aldrich, Sydney, Australia) and sterile water. The pumps were filled with 200μl of AVE0991 solution and incubated in saline overnight at 37°C were implanted in 14 rats in the peritoneal cavity during BDL surgery and delivered 28μg/kg/h for 2 weeks as previously described [2 (link)]. Twelve BDL rats were implanted with pumps containing vehicle alone and served as BDL control group. Sixteen sham-operated rats were also included. Animals were sacrified and livers analyzed 2 weeks after BDL and AVE0991 administration.
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2

Reagents for Cell Signaling Experiments

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AVE0991 was a kind gift from Sanofi‐Aventis (Frankfurt, Germany); A799 was from Bachem (Bubendorf, Switzerland); IFN‐γ and TNF‐α were supplied by PeproTech EC (London, UK); insulin was from Cell Applications (San Diego, CA) and PGF was supplied by Cayman Chemical Company (Ann Arbor, MI). Sigma‐Aldrich (St Louis, MO) supplied ACh, LPS (from Escherichia coli 0111:B4;), dexamethasone, IBMX and SNP.
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3

Reagents and Materials Sourcing

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All chemicals used in this study were purchased from Sigma (St. Louis, MO) except A779 (Bachem, Torrance, CA), PD123319, CGP42112, and HOE140 (Tocris, Ellisville, MO), and DETA NONOate (Cayman, Ann Arbor, MI). AVE0991 was a generous gift from Sanofi-Aventis (Frankfurt, Germany).
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4

Transaortic Constriction and Pharmacological Interventions in Mice

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The authors declare that all supporting data are available within the article and its online supplementary files.
All experimental procedures were designed in accordance with the National Institutes of Health guidelines and approved by the Animal Welfare Committee and the Center for Laboratory Animal Medicine and Care in the University of Texas Health Science Center at Houston. Ten to eleven-week-old male C57BL/6J wild-type mice were purchased from Jackson Laboratory, Bar Harbor, ME, USA. At the age of 12 weeks, mice weighing 23-31grams (27.5±1.9g) were randomized to treatment with or without various drugs in the drinking water for three days, then underwent TAC or sham operation. Treatment was continued for two weeks post-operation. Losartan (sc-204796A, Santa Cruz Biotechnology, 0.6g/L)12 (link), captopril (sc-200566A, Santa Cruz Biotechnology, 75mg/L)11 , AT2R agonist C21 (a gift from Vicore Pharma, 300μg/kg/day),25 (link) AT2R antagonist PD123319 (P186-10MG, Sigma-Aldrich, 3mg/kg/day)20 (link) and MasR agonist AVE0991 (a gift from Sanofi-Aventis, Frankfurt/Main, 576μg/kg/day)26 (link) were administrated 1 hour prior operation to two weeks post-operation via intraperitoneal injection. An illustration of the signaling pathways and the drug targets are presented in Supplemental figure 1A.
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5

Glucose Tolerance in Zucker Fatty Rats

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Male Zucker fatty rats (fa/fa) (n = 11) were purchased from Harlan (Udine, Italy). The animals were housed in a 12-hour light/dark cycle with access to water and standard diet ad libitum. Animals were separated to two groups. The control group was treated with vehicle (30% solution of cyclodextrin), and the experimental group received AVE0991 (Sanofi-Aventis, Frankfurt, Germany) (0.5 mg/kg BW/day in 30% solution of cyclodextrin) via osmotic minipumps (ALZET, CA, USA) for two weeks. The intraperitoneal glucose tolerance test (IPGTT) was performed to assess glucose clearance. On the 12th day, overnight-fasted animals were administered an i.p. injection of dextrose solution at a dose of 2 g/kg body weight. Glycaemia was measured in the tail vein blood immediately and in 30-minute intervals for 2 hours after glucose administration using a glucometer (Accu-Chek Active, Roche Diagnostics, Switzerland). After 2 days of recovery, overnight-fasted animals were sacrificed by decapitation at the age of 8 months. Experimental procedures involving animals were approved by the Jagiellonian University Ethical Committee on Animal Experiments.
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