Losartan potassium

A-779 (5-L-isoleucine-7-D-alanine-1-7-angiotensin II, trifluoroacetate salt); AM251 [(N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide)]; angiotensin 1–7; angiotensin II (Tocris Bioscience, Bristol, UK); betadine (Egis Pharmaceuticals PLC, Budapest, Hungary); buprenorphine (Richter Pharma AG, Wels, Austria); CP55940 [(−)-cis-3-[2-hydroxy4-(1,1-dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)cyclohexanol] (Sigma Aldrich, St. Louis, MO, USA); losartan potassium (Tocris Bioscience, Bristol, UK); paracetamol (Sequoia, Warsaw, Poland); PD123319 ((6S)-1-[[4-(dimethylamino)-3-methylphenyl]methyl]5-(2,2-diphenylacetyl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylic acid, di(2,2,2-trifluoroacetate)) (Sigma Aldrich, St. Louis, MO, USA); pentobarbitone sodium (Biowet, Puławy, Poland).
Drugs were dissolved in saline with the following exceptions. AM251 was dissolved in a mixture of ethanol, Cremophor El, DMSO and saline (1:1:1:9.5) for i.v. injections and in DMSO and saline (1:9) for PVN injections. CP55940 was dissolved in a 19% solution of cyclodextrin. None of the used vehicles significantly altered any of the cardiovascular parameters by itself (data not shown).

The rat neonatal H9c2 cardiac myoblast cell line was obtained from the Korea Cell Line Bank (Seoul, Korea). The cells were cultured in Dulbecco’s modified Eagles medium (DMEM)/high glucose with 10% fetal bovine serum and 1% penicillin-streptomycin (Corning/Mediatech, Inc., Manassas, VA, USA,).
DOX was purchased from Tocris Bioscience (Cat#2252, Bristol, UK). LY294002 (Cat#1130), the PI3K inhibitors, cilengitide (Cat#5870), and losartan potassium (Cat#3798), the integrin inhibitors α_vβ₃ and α_vβ₅, and the angiotensin II type 1 receptor (AT1R) inhibitors respectively, were purchased from Tocris Bioscience. The ILK inhibitor Cpd22 (Cat#407331) was obtained from EMD Millipore Corp. (Darmstadt, Germany). The survivin inhibitor YM-155 was purchased from MedChem Express (Cat#HY-10194, Monmouth Junction, NJ, USA).

Angiotensin II (Ang II; Tocris Cat# 1158), Ang-(1–7) (Tocris Cat# 1562), AT2R-Agonist CGP42112 (Tocris Cat# 2569), Losartan potassium (Tocris Cat# 3798) and (+-)-Norepinephrine (+)-bitartrate salt (Sigma Cat# A0937, Cas # 3414-63-9) were dissolved in sterile H₂O. Ang II and CGP 42112 were used at a final working concentration of 500 nM, Ang-(1–7) at 10 nM, losartan at 5 μM and norepinephrine at 10 μM.
Regarding all pharmacological LFP experiments with Ang II, Ang-(1–7), AT2R-Agonist and losartan, rhythmic activity was recorded for 15 min under control condition. Treatment was applied for 20 min followed by a 40 min washout (if performed), with one exception. In one experiment losartan was applied for 10 min after which Ang II was co-applied for another 20 min. Regarding the calcium imaging experiments, calcium activity was recorded under control condition for 30–60 s after which Ang II was applied and recorded for another ∼7 min. Immediately after this, a new recording was started in which the calcium activity under the previously applied Ang II was recorded again for 30–60 s, then norepinephrine was co-applied.