Olmesartan

Male C57BL/6J mice, 7–10 weeks old, were purchased from CLEA Japan (Tokyo, Japan). To generate the SSHTN model, mice were administered l-NAME (0.5 mg/mL; Sigma-Aldrich, St. Louis, MO, United States) in their drinking water for 2 weeks to inhibit nitric oxide synthesis, followed by a 2-weeks washout period, and a 3-weeks exposure to a high-salt diet (CE-2 containing 4% NaCl; CLEA Japan) (Lopez Gelston et al., 2018 (link)) (Figure 1A). Normal control mice received tap water and standard diet for 7 weeks. All diets and water were provided ad libitum. Systolic blood pressure (SBP) was monitored weekly by tail-cuff plethysmography using a blood pressure monitor (MK-1030; Muromachi Kikai Co., Tokyo, Japan) in previously trained mice. All animal care and experiments were approved by Tohoku University of Science Animal Care and Use Committee. LPS from Escherichia coli, angiotensin II, and olmesartan (angiotensin II type 1 receptor blocker) were purchased from Sigma-Aldrich. Recombinant murine TNF-α was prepared in our laboratory as described previously (Kitaura et al., 2004 (link)).

IL-1β and IL-10 cytokines were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany) and resuspended in sterile water. TNFα was purchased from Peprotech (Rocky Hill, NJ, USA) and resuspended in sterile water. Olmesartan, losartan, and enalapril were purchased from Sigma-Aldrich and dissolved in sterile water. Metformin was purchased by Sigma-Aldrich and dissolved in sterile water.

Captopril, Olmesartan, CGP42112A, and alloxan monohydrate were purchased from Sigma Chemical Co. (Saint Louis, MO, USA). PD123319 from Cayman Chemical Co. (Ann Arbor, MI, USA). Ethanol, methanol, and xylene from Merck (Rio de Janeiro, RJ, Brazil). Sodium heparin and sterile saline solution from Roche (São Paulo, SP, Brazil). All solutions were prepared immediately before use.