L phenylephrine hydrochloride

The chemicals used in the present study are as follows: acetylcholine chloride (Daiichi Pharmaceutical Co., Ltd., Chuo-ku, Tokyo, Japan), l-phenylephrine hydrochloride and NAC (Sigma-Aldrich Co., LLC., St. Louis, MO, USA), vaspin (mouse) and visfatin (rat) (cat. no. AG-40A-0094 and AG-40A-0058, Adipogen Corp., San Diego, CA, USA), omentin (rat) (cat. no. ALX-522-132, Enzo Life Sci., Inc., Farmingdale, NY, USA), and apelin-12 (human, rat, mouse, bovine) (cat. no. 057-23, Phoenix Pharmaceuticals, Inc., Burlingame, CA, USA). Other chemicals of analytical reagent grade were purchased from NACalai Tesque Inc. (Kyoto, Kyoto, Japan). Stock solutions of test-compounds were prepared in distilled water.

L-phenylephrine hydrochloride, ACh and sodium pentobarbital were purchased from Sigma-Aldrich (St. Louis, USA). Salts and reagents used were of analytical grade from Sigma-Aldrich and Merck (Darmstadt, Germany).

The drugs used were obtained from the following sources: L-phenylephrine hydrochloride, acetylcholine chloride (Sigma-Aldrich, St. Louis, MO, USA), native human thrombospondin 1 (Novus Biologicals, Littleton, CO, USA; Athens Research & Technology, Athens, GA, USA; Thermo Fisher Scientific, Anthem, AZ, USA), recombinant human thrombospondin 1 (R&D Systems, Minneapolis, MN, USA; BioVision, Milpitas, CA, USA), recombinant human CD47 peptide (Bon Opus, previously NovoProtein, Millburn, NJ; Abcam, Cambridge, MA, USA). All drugs were dissolved and diluted in physiological salt solution. All animal studies were performed under a protocol approved (16 August 2018) by the Midwestern University Institutional Animal Care and Use Committee (IACUC) in accordance with NIH guidelines. Male C57BL/6 mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Hydroxylamine hydrochloride, pyridine, magnesium sulphate, l-phenylephrine hydrochloride (PE), acetylcholine chloride (ACh), N^G-nitro-l-arginine methyl ester (l-NAME) were bought from Sigma-Aldrich (St Luis, MO, USA). The metabolite and oxime were dissolved in DMSO (0.1% final concentration).

Silica gel GF254 analytical chromatoplates, Silica gel grade 60 A for column chromatography were purchased from Merck Co., Santiago, Chile. Precoated thin-layer chromatography TLC plates SIL G-100 UV254, 1.0 mm, preparative were purchased from Machery−Nagel (GmbH&Co, KG), Dueren, Germany. 8-oxo-9-dihydromakomakine was 99% purified through recrystallization.
The drugs used were l-phenylephrine hydrochloride (PE), N^ω-nitro-l-arginine methyl ester (l-NAME), ouabain, and Bay K8644 (Sigma-Aldrich, St Luis, MO, USA). Caffeine, Tetraethylammonium (TEA), and Barium chloride dihydrate (BaCl₂) were obtained from Merck (Darmstadt, Germany). The drugs were dissolved in distilled deionized water (deionized water Millipore) and kept at 4 °C. The stock solution of 8-oxo-9-dihydromakomakine was dissolved in DMSO (Merck, Germany). Final DMSO concentration in the organ bath was lower than 0.1%. The Krebs-Ringer bicarbonate (KRB) solutions were freshly prepared before each experiment.

l-Phenylephrine hydrochloride, ACh chloride, SNP, urethane, OUA, L-NAME, TEA, 4-AP, IbTX, ChTX and apamin were purchased from Sigma-Aldrich (St. Louis, USA). The salts and reagents used were of analytical grade from Sigma-Aldrich and Merck (Darmstadt, Germany).