Losartan concentration of 0.6 grams/liter was prepared to give an estimated Losartan dose of 50–70 mg/kg/day in the drinking water.[21 (link), 22 ] 650 mg of Losartan tablets were dissolved in 1080 ml of water to achieve a concentration of 0.6 g/L. The mixture was then double-filtered using a vacuum filtration system to remove the remaining excess sediment.[23 ] For cell culture purposes, we used 10 μM Losartan (Cayman Chemicals, Ann Arbor, Michigan, USA) for 48 hours.[24 (link), 25 (link)]
Losartan
Manufactured by Cayman Chemical
Sourced in United States
Losartan is a synthetic compound that functions as an angiotensin II receptor antagonist. It is commonly used as a reference standard in analytical and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Telmisartan, by Cayman Chemical (3 mentions)
Gw9662, by Merck Group (3 mentions)
Trypsin edta solution, by Thermo Fisher Scientific (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Plasticware for cell culture, by Thermo Fisher Scientific (2 mentions)
P ampk thr172, by Cell Signaling Technology (2 mentions)
D glucose, by Merck Group (2 mentions)
D mannitol, by Merck Group (1 mentions)
P enos thr497, by BD (1 mentions)
Minimum essential medium (mem), by Welgene (1 mentions)
Dulbecco s phosphate buffered saline dpbs, by Welgene (1 mentions)
N 1 naphthyl ethylenediamine, by Merck Group (1 mentions)
Okadaic acid, by Merck Group (1 mentions)
Valsartan, by Merck Group (1 mentions)
Rosiglitazone, by GlaxoSmithKline (1 mentions)
Verapamil, by Thermo Fisher Scientific (1 mentions)
Cefadroxil, by Merck Group (1 mentions)
Digoxin, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated goat anti rabbit igg, by Cell Signaling Technology (1 mentions)
4 hydroxy tempo, by Merck Group (1 mentions)
10 protocols using losartan
1
Preparation and Use of Losartan Solution
2
Regulation of Endothelial Nitric Oxide Synthase by Angiotensin II Receptor Antagonists
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Telmisartan and losartan were purchased from Cayman Chemicals (Ann Arbor, MI, USA). Fimasartan was a gift from Boryung Pharmaceuticals (Seoul, Korea). D-glucose, D-mannitol, GW9662, acetylcholine (ACh), phenylephrine (PE), sulfanilamide, N-(1-Naphthyl)ethylenediamine, okadaic acid, and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against eNOS, p-eNOS-Ser1179, p-eNOS-Thr497, and PP2Ac were purchased from BD Transduction Laboratories (Lexington, KY, USA). Antibodies against Akt, p-Akt-Ser473, AMPK, p-AMPK-Thr172 were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibodies against p-eNOS-Ser116 and β-actin were purchased from Abcam (Cambridge, MA, USA) and Sigma-Aldrich, respectively. Minimum essential medium (MEM) and Dulbecco's phosphate-buffered saline (DPBS) were obtained from Welgene Inc. (Gyeongsan, Korea). Fetal bovine serum (FBS), penicillin and streptomycin antibiotics, trypsin–EDTA solution, and plasticware for cell culture were purchased from Gibco-BRL (Gaithersburg, MD, USA). All other chemicals used were of the purest analytical grade available.
3
Sourcing and Characterizing Pharmaceutical Compounds
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Digoxin, 95% and (+/−)-verapamil, 99% were obtained from Acros Organics (Morris Plains, NJ, USA); indomethacin, 98% and ranitidine hydrochloride were purchased from Alfa Aesar GmbH and Co KG (Karlsruhe, Germany), whereas losartan was obtained from Cayman Chemical (Ann Arbor, MI, USA). Atropine sodium salt, cefadroxil, fluvastatin natrium and valsartan were purchased from Sigma-Aldrich (St. Louis, MO, USA); rosuvastatin calcium salt was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Fexofenadine and sulfasalazine were purchased from Fluorochem Ltd. (Hadfield, UK), and rosiglitazone was obtained from GlaxoSmithKline (Brentford, UK). Agar, calcium chloride dihydrate, glucose hydrate, magnesium chloride hexahydrate, potassium chloride, sodium chloride, sodium phosphate monobasic and sodium hydrogen carbonate were obtained from Hänseler AG (Herisau, Switzerland).
4
Pharmacological Modulation of Endothelial Function
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Potassium chloride (KCl), PE, Nω-nitro-L-arginine methyl ester (L-NAME, a NOS inhibitor), ACh, sodium nitroprusside (SNP), 4-Hydroxy-TEMPO (tempol, a superoxide anion scavenger), indomethacin (a non-selective cyclooxygenase (COX) inhibitor), and methyl-β-cyclodextrin (CD, a cholesterol depleter which disassembles caveolae) were purchased from Sigma-Aldrich (St. Louis, MO, USA); losartan (an Ang II type 1 receptor antagonist) was from Cayman Chemical Company (Ann Arbor, MI, USA). Drug concentrations were expressed as final molar concentrations in the bath chamber. 10× radio-immunoprecipitation assay (RIPA) lysis buffer was from Millipore Corporate Headquarters (Temecula, CA, USA); bicinchoninic acid (BCA) protein assay kit and Pierce enhanced chemiluminescent (ECL) Western Blotting Substrate were from Thermo Fisher Scientific (Rockford, IL, USA); anti-p-eNOSThr495 was from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-p-eNOSSer1177 was from Abcam plc (Cambridge, UK); anti-eNOS, anti-adenosine monophosphate-activated protein kinase (anti-AMPK), anti-Cav-1, anti-β-actin, and horseradish peroxidase (HRP) conjugated goat anti-mouse IgG were from BD Transduction Laboratorie (Lexington, KY, USA); HRP conjugated goat anti-rabbit IgG was from Cell Signaling Technology (Danvers, MA, USA).
5
Molecular Mechanisms of Insulin Signaling
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Telmisartan and losartan were purchased from Cayman Chemical (Ann Arbor, MI, USA). Fimasartan was a kind gift from Boryung Pharmaceutical (Seoul, Korea). D-Glucose, GW9662, sodium lactate, HEPES, dimethyl sulfoxide (DMSO), and bovine insulin were obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against Akt, p-Akt-Ser473, GSK3β, p-IRS-1-Ser302, p-IRS-Ser307, p-IRS-Ser318, p-IRS-Ser1101, p-IRS-Tyr632, p-IRS-Tyr896, IRS-1, and actin were purchased from Cell Signaling Technology (Boston, MA, USA). Antibodies against glucose-6-phosphatase α (G6Pase-α), phosphoenolpyruvate carboxykinase (PEPCK), and PKCζ, p-PKCζ-Thr410 were purchased from Santa Cruz Biotechnology (La Jolla, CA, USA). Antibodies against p-GSK3β-Ser9 and hemagglutinin (HA) were obtained from BD Biosciences (San Jose, CA, USA) and Covance Inc. (Emeryville, CA, USA), respectively. Low or high glucose Dulbecco’s modified Eagle’s medium (DMEM) and Dulbecco’s phosphate-buffered saline were purchased from Welgene Inc. (Gyeongsan, Korea). Sodium pyruvate, fetal bovine serum (FBS), penicillin and streptomycin antibiotics, L-glutamine, and trypsin-EDTA solution were purchased from Gibco-BRL (Gaithersburg, MD, USA). Plasticware for cell culture was purchased from Corning Inc. (Oneonta, NY, USA) or SPL Life Sciences (Pocheon, Korea). All other chemicals were of the purest analytical grade.
6
Inhibition of Cellular Signaling Pathways
7
Aortic Relaxation Mechanisms Investigation
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TMP was purchased from the National Institutes for Food and Drug Control (Beijing, China). Phenylephrine (Phe), acetylcholine (ACh), apocynin, diphenyleneiodonium (DPI), vitamin E, catalase, Tiron (4,5-dihydroxy-1,3-benzene-disulfonic acid), and diethyldithiocarbamic acid (DETCA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dihydroethidium (DHE) was purchased from Molecular Probes (Eugene, OR, USA). Losartan was purchased from Cayman Chemical Co. (Ann Arbor, MI, USA). Apocynin and Losartan were dissolved in dimethyl sulfoxide (DMSO), and the other compounds were dissolved in distilled water. The DMSO vehicle did not affect ACh-induced aortic ring relaxation at the concentration employed in this study (0.1% v/v).
8
Angiotensin II Dose-Response on AFC
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To investigate the effect of angiotensin II on AFC, we established a dose response curve of Ang II perfused through the pulmonary circulation and instilled to the lungs: 10−10, 10−9, 10−8, 10−7 and 10-6M. While 10−9 M Ouabain (Cat# 1076, Tocris Biosiences) and 10-6M Amiloride (Cat# A7410, Sigma Aldrich) were given through the tail vein 30 minutes prior to sacrificing the rat, 10−6 M Norepinephrine (Cat# 1468501, Sigma Aldrich) was perfused through the pulmonary circulation of the isolated lungs. Losartan (0.02mg/gr rat/day) was added to drink water for three days prior to sacrificing the rat (Cat# 10006594, Cayman chemical). PD123319 was given through the tail vein 30 minutes prior to sacrificing the rat (100μg/kg rat), and perfused through the pulmonary circulation (10μg/ml) since it has a short half time (22 minutes) (Cat# P186, Sigma Aldrich).
9
Inhibition of PDGF-induced signaling
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Telmisartan, losartan, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Cayman Chemicals (Ann Arbor, MI, USA). Fimasartan was a kind gift from Boryung Pharmaceuticals (Seoul, Korea). GW9662 and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Compound C was purchased from Calbiochem (Darmstadt, Germany). Recombinant rat platelet-derived growth factor (PDGF)-BB protein was purchased from Novus Biologicals (Centennial, CO, USA). Antibodies against mammalian target of rapamycin (mTOR), p-mTOR-Ser2448, p70 S6 kinase (p70S6K), p-p70S6K-Thr389, AMPK, p-AMPK-Thr172, acetyl-CoA carboxylase (ACC), and p-ACC-Ser79 were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibody against β-actin was purchased from Sigma-Aldrich. Dulbecco's modified Eagle's medium (DMEM) was obtained from Fisher Scientific (Ottawa, Canada). Dulbecco's phosphate-buffered saline (DPBS), fetal bovine serum (FBS), penicillin and streptomycin antibiotics, trypsin–EDTA solution, and plasticware for cell culture were purchased from Gibco-BRL (Gaithersburg, MD, USA). All other chemicals used were of the purest analytical grade available.
10
Pharmacological Modulation of Vascular Reactivity
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Acetylcholine (Ach), phenylephrine (Phe), Indomethacin (Indo), and angiotensin II (Ang II) were bought from Sigma-Aldrich (St. Louis, MO, United States). 9,11-dideoxy-9α,11α-methanoepoxy Prosta-glandin F2α (U46619), L-NAME, nifedipine, 1H-[1,2,4]-oxadiazolo-[4,3-alpha]-quinoxalin-1-one (ODQ) and Losartan, Enalapril, PD123319 were from Cayman Chemical (MI, United States). Fura-2 was supplied by Beyotime Biotechnology (Shanghai, China). THP hydrochloride (purity by HPLC ≥98.0%) was bought from Chengdu Must Bio-Technology Co., Ltd. (Chengdu, China). THP was dissolved in dimethyl sulfoxide (DMSO) while other drugs were prepared in distilled water.
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