The study aims to explore the association between black cumin extract supplementation to endothelial dysfunction markers and aortic Intima Media Thickness (IMT) in rats with subchronical cigarette smoke exposure. Fifty Wistar rats (Rattus norvegicus) were classified into five kind of groups: negative control group (NC); positive control group (PC), which was shown up to 40 cigarettes/day for about 4 weeks; and three groups shown up to cigarette smoke and it has administration of extract of black cumin ethanolic for about four weeks but in different amount of doses: 0.3 g/kg/day (T1); 0.6 g/kg/day (T2); and 1.2 g/kg/day (T3). After getting intervented, aorta was examined to measure the level of vascular cell adhesion molecule-1 (VCAM-1), endothelial nitric oxide synthase (eNOS), and also IMT. The dominant active substances of black cumin are linoleic acid (71.31%), palmitate acid (15.86%), stearate (4.69%), octadicadienal (3.8%), beta monolinolein (1.8%), and thymoquinone (0.28%). eNOS level significantly increased in T1 compared to PC with p=0.001. A significant descending in VCAM-1 was found in T2 (p=0.014) and T3 (p=0.044) compared to PC. Aortic IMT decreased significantly in T1 (p=0.009), T2 (p=0.000), and also T3 (p=0.043) than the PC. Increased eNOS, decreased VCAM-1 and IMT following black cumin extract supplementation showed that black cumin may prevent endothelial dysfunction. [ABSTRACT FROM AUTHOR]