The eNOS Glu298Asp (rs1799983) polymorphism of the NOS3 gene has been implicated as a risk factor for cardiovascular diseases; however, not all studies find significant associations. We examined this possibility in a Russian (Siberian) population. One hundred patients with chronic heart failure and 40 controls were investigated. PCR analysis was performed on DNA samples. The aim was to evaluate a possible association between the (Glu298Asp) polymorphism (rs1799983) of the NOS3 gene and susceptibility to chronic heart failure in the Russian population. We evaluated genotype distributions in patient and control groups and assessed the relationship between genotypes and chronic heart failure. We found that this polymorphism is not associated with increased risk of chronic heart failure in our study cohort. In conclusion, testing of the NOS3 gene polymorphism does not seem useful for evaluating predisposition for chronic heart failure or its diagnosis and prognosis.