Glibenclamide

Vasoactive signaling pathways were inhibited mice with the following protocols that have been previously validated. Inhibition of eNOS was performed with 75 μg/kg L-nitroarginine methyl ester (L-NAME, Santa Cruz Biotech., Santa Cruz, CA) given IP 30 min prior to study. Blockade of adenosine A₂-receptors was performed with 50 μg/g of 4-(2-[7-Amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM-241385, Abcam PLC, Cambridge, MA) given IV 1 hr prior to study. This dose is predicted to inhibit both A_2a and A_2b receptors. Accelerated metabolism of ATP was performed with 4.0 U/g apyrase (Sigma-Aldrich, St. Louis, MO) given IP 1 hour prior to study; while inhibition of ectonucleotidases involved in ATP metabolism was performed by 1 μg/g of sodium polytungstate (POM-1, VWR, Radnor, PA) given by IP injection 30 min prior to study. For inhibition of the K_ATP receptor was performed by 15 mg/kg glibenclamide (Santa Cruz Biotechnology, Dallas, TX) given IV 30 min prior to study. For inhibiting intracellular signaling pathways, L-NAME (20 mg/kg) was administered by IV route 10 min prior to imaging; and indomethacin (2 mg/kg) was given by IP route 24 hrs and again 1 hr prior to study.