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6 protocols using phenylephrine phe

1

Isolated Vascular Tissue Preparation

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Calcium chloride dihydrate (CaCl2·2H2O), potassium chloride (KCl), and sodium bicarbonate (NaHCO3) were purchased from Vetec (Rio de Janeiro, RJ, Brazil). Glucose (C6H12O6), magnesium sulfate heptahydrate (MgSO4·7H2O), hydrochloric acid (HCl), and monobasic potassium phosphate (KH2PO4) were from Nuclear (Porto Alegre, RS, Brazil). Sodium chloride (NaCl) was purchased from Dinâmica (Diadema, SP, Brazil) and acetylcholine chloride (ACh) from Merck (Brazil). Phenylephrine (PHE) was obtained from Pfizer (USA), and Nω-nitro-L-arginine methyl ester (L-NAME) was purchased from Sigma-Aldrich (Brazil). Ethylenediamine tetraacetic acid (EDTA) (1 : 250) came from BioTécnica-Advanced Biotechnology (Brazil), and carbogen mixture (95% O2 and 5% CO2) was obtained from White Martins (Brazil). All substances were weighed on an analytical balance, GEHAKA model AG 200 (Sao Paulo, SP, Brazil).
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2

Vasoactive Compound Preparation Protocol

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Magnesium sulfate (MgSO4), potassium chloride (KCl), calcium chloride (CaCl2), sodium chloride (NaCl) and formaldehyde were purchased from Vetec Química Fina Ltda. (Brazil). Glucose (C6H12O6) and sodium bicarbonate (NaHCO3) were purchased from Dinâmica (Brazil). Potassium monobasic phosphate (KH2PO4), sodium hydroxide (NaOH) and hydrochloric acid (HCl) were purchased from Nuclear (Brazil). These substances, except glucose, NaCl and NaHCO3 were diluted in distilled water to obtain each solution, which were maintained under refrigeration.
Phenylephrine (Phe) was purchased from Pfizer (USA). Acetylcholine (ACh), sodium nitroprusside (SNP), L-Nω-nitroarginine methyl ester (L-NAME), indomethacin, tempol, apocynin, Cremophor® and R-(-)-apomorphine were obtained from Sigma-Aldrich (Brazil). All substances were diluted in distilled water as needed for each experimental protocol. The carbogen mixture (95% O2 and 5% CO2) was acquired from White Martins (Brazil).
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3

Vascular Reactivity Assay Reagents

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Calcium chloride dihydrate (CaCl2·2H2O), magnesium sulfate heptahydrate (MgSO4·7H2O), and glucose (C6H12O6) were purchased from Vetec (Rio de Janeiro, RJ, Brazil). Sodium bicarbonate (NaHCO3), sodium chloride (NaCl), and potassium chloride (KCl) were purchased from Nuclear (Porto Alegre, RS, Brazil). Monobasic potassium phosphate (KH2PO4), monobasic sodium phosphate (NaH2PO4), sodium hydroxide (NaOH), and hydrochloric acid (HCl) were purchased from Dinâmica (Diadema, SP, Brazil).
Phenylephrine (PHE) was purchased from Pfizer (USA). ACh, R-(-)-apomorphine, Nω-nitro-L-arginine methyl ester (L-NAME), indomethacin, tempol, apocinin, ethylenediamine tetraacetic acid (EDTA), MDA, and 1,1-diPHEnyl-2-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich (Brazil). To obtain the stock solutions, substances used in the functional experiments were dissolved and diluted in distilled water (indomethacin and apocinin were dissolved in absolute alcohol 96° GL), being, respectively, kept at 4 or -20°C. The carbogen mixture (95% O2 and 5% CO2) was obtained from White Martins (Brazil). All substances were weighed on an analytical balance, GEHAKA model AG 200 (Sao Paulo, SP, Brazil).
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4

Evaluation of Pharmacological Agents

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The R-(-)-apomorphine, malondialdehyde (MDA), 1,1-diphenyl-2-picryl hydrazyl (DPPH) and acetylcholine (ACh) were obtained from Sigma-Aldrich (São Paulo, Brazil). Phenylephrine (Phe) was purchased from Pfizer (Kalamazoo, USA). Magnesium sulphate (MgSO4), potassium chloride (KCl), calcium chloride (CaCl2), sodium chloride (NaCl) and formaldehyde were purchased from Vetec Química Fina Ltda. (São Paulo, Brazil). Glucose (C6H12O6) and sodium bicarbonate (NaHCO3) were purchased from Dinâmica (São Paulo, Brazil). Potassium monobasic phosphate (KH2PO4), sodium hydroxide (NaOH) and hydrochloric acid (HCl) were purchased from Nuclear (São Paulo, Brazil). These substances, except the glucose, NaCl and NaHCO3 were diluted in distilled water to obtain each solution, which were maintained under refrigeration.
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5

Balanced Salt Solution Preparation

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Glucose (C6H12O6), monobasic potassium phosphate (KH2PO4), hydrochloric acid (HCl), and magnesium sulfate heptahydrate (MgSO4.7H2O) were obtained from Nuclear (Brazil). Sodium chloride (NaCl) was purchased from Dinâmica (Brazil). Potassium chloride (KCl), calcium chloride dihydrate (CaCl2.2H2O), and sodium bicarbonate (NaHCO3) were acquired from Vetec (Brazil) and acetylcholine chloride (ACh) from Merck (Brazil). Phenylephrine (PHE) was obtained from Pfizer (United States) and aminophylline and indomethacin from Sigma Aldrich (Brazil). Ethylenediaminetetraacetic acid (EDTA) (1:250) was purchased from BioTécnica-Advanced Biotechnology (Brazil) and carbogen mixture (95% O2 and 5% CO2) from White Martins (Brazil). The weighing of the substances was done with a GEHAKA analytical balance model AG 200 (Brazil).
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6

Preparation of Pharmacological Solutions

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Calcium chloride dihydrate (CaCl2.2H2O), potassium chloride (KCl) and sodium bicarbonate (NaHCO3) were purchased from Vetec (Rio de Janeiro - RJ, Brazil). Glucose (C6H12O6), magnesium sulfate heptahydrate, (MgSO4.7H2O), hydrochloric acid (HCl) and monobasic potassium phosphate (KH2PO4) were obtained from Nuclear (Porto Alegre - RS, Brazil). Sodium chloride (NaCl) was purchased from Dynamics (Diadema-SP, Brazil). The acetylcholine (ACh) was obtained from Merck (Brazil). Phenylephrine (PHE) was acquired from Pfizer (USA). Nω-nitro-L-arginine methyl ester (L-NAME) was purchased from Sigma-Aldrich (Brazil). The ethylenediaminetetraacetic acid (EDTA) (1:250) was purchased from BioTécnica-Advanced Biotechnology (Brazil). The carbogen mixture (95% O2 and 5% CO2) was obtained from White Martins (Brazil). All substances were weighed on an analytical balance, model AG 200 GEHAKA (São Paulo-SP, Brazil).
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