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Bekesho Geleta,1,2 Eyasu Makonnen,2 Asfaw Debella,1 Abiy Abebe,1 Netsanet Fekadu,1 1Directorate of Traditional and Modern Medicine Research, Ethiopian Public Health Institute, 2Department of Pharmacology, School of Medicine, College of Health Sciences, Addis Ababa University, Addis Ababa, Ethiopia Abstract: Moringa stenopetala, a plant belonging to the family of Moringaceae, is traditionally used for the treatment of hypertension and diabetes in Ethiopia. This study evaluates the in vitro vasodilatory effect of the extract of M. stenopetala leaves and the possible mechanisms in precontracted isolated thoracic aorta of guinea pigs. A guinea pig was sacrificed by gentle cervical dislocation, and the thoracic aortic ring was removed, cut spirally, and mounted in an organ bath containing Krebs–Henseleit physiological solution maintained at 37°C, and then the solution was aerated with carbogen (95% O2 and 5% CO2). The vasodilatory activity of cumulative doses of M. stenopetala extracts and fractions was evaluated on intact and denuded endothelium of isolated whole, spirally cut thoracic aortic strips of guinea pigs precontracted with potassium chloride (80 mM), epinephrine (1 µM), methylene blue (10 µM), and glibenclamide (10 µM) using polygraph. All extracts showed a relaxant effect in precontracted isolated whole, spirally cut thoracic aortic strips of guinea pigs in a dose-dependent manner, whereas the greater percentage of relaxant effect was shown with the addition of crude extracts in 80 mM of potassium chloride (99.10% and 95.56% for ethanol and aqueous crude extracts, respectively), and 1 µM of epinephrine (82.85% and 90.16% for ethanol and aqueous crude extracts, respectively) in precontracted isolated whole, spirally cut thoracic aortic strips of guinea pigs. Hence, the possible mechanism of relaxation might be mediated through the blockade of receptor-operated calcium influx and L-type voltage-dependent calcium channels. The aqueous extract showed more significant in vitro vasodilatory effect than its fractions and 70% ethanol extract. Keywords: in vitro vasodilatory, Moringa stenopetala, aortic strips, organ bath, mechanism of action, guinea pigs