Nitric oxide (NO) is known to be one of the most important regulatory compounds within the cardiovascular system where it is central for functions such as regulation of blood pressure, blood flow, and vascular growth. The bioavailability of NO is determined by a balance between, on one hand, the extent of enzymatic and non-enzymatic formation of NO and on the other hand, removal of NO, which in part is dependent on the reaction of NO with reactive oxygen species (ROS). The presence of ROS is dependent on the extent of ROS formation via mitochondria and/or enzymes such as NAD(P)H oxidase (NOX) and xanthine oxidase (XO) and the degree of ROS removal through the antioxidant defense system or other reactions. The development of cardiovascular disease has been proposed to be closely related to a reduced bioavailability of NO in parallel with an increased presence of ROS. Excessive levels of ROS not only lower the bioavailability of NO but may also cause cellular damage in the cardiovascular system. Physical activity has been shown to greatly improve cardiovascular function, in part through improved bioavailability of NO, enhanced endogenous antioxidant defense and a lowering of the expression of ROS-forming enzymes. Regular physical activity is therefore likely to be a highly useful tool in the treatment of cardiovascular disease. Future studies should focus on which form of exercise may be most optimal for enhancing NO bioavailability and improving cardiovascular health.