Abstract. The present study aimed to investigate the changes of Rho-associated kinase (Rho-kinase) and neuronal nitric oxide synthase (nNOS) expression in age-associated erectile dysfunction (ED) in Sprague-Dawley (SD) rats. A total of 100 intact male SD rats were divided into 20 groups according to their age (5-24 months; rats that were the same age in months were in the same group). Erectile response measurements were performed and the functional index intracavernosal pressure/mean arterial pressure (ICP/MAP) was tested, followed by detection of Rho-kinase and nNOS protein by western blot analysis. Finally, correlation analyses of the association between ICP/MAP and Rho-kinase, nNOS, or nNOS/Rho-kinase, as well as between age and nNOS or Rho-kinase, were performed. The functional index ICP/MAP decreased with age in SD rats. Moreover, the expression of nNOS protein decreased, while Rho-kinase expression increased, indicating that the nNOS/Rho-kinase ratio decreased with age. The Pearson's correlation coefficients for the association between ICP/MAP and Rho-kinase, nNOS and nNOS/Rho-kinase ratio were -0.917, 0.853 and 0.937, respectively (P<0.01). Furthermore, nNOS was found to be significantly negatively correlated with age (r=-0.855; P<0.01), whereas Rho-kinase was positively correlated with age (r=0.943; P<0.01). Age-associated ED was therefore correlated with decreased nNOS and increased Rho-kinase, indicating that the nNOS/Rho-kinase ratio may be used as a candidate indicator of age-associated ED.