Symmetric dimethylarginine (SDMA) is a serum biomarker of renal damage in dogs. Moreover, SDMA concentration is an independent predictor of development of severe heart failure (HF) in humans with cardiac disease. This study evaluates whether the serum concentration of SDMA in dogs with myxomatous mitral valve disease (MMVD) is influenced by the severity of heart disease, pulmonary hypertension (PH) and treatment of HF. A total of 99 client-owned dogs were included in this retrospective case-control study; 78 dogs were affected by MMVD and classified according to the American College of Veterinary Internal Medicine (ACVIM) guidelines, and 21 were healthy controls. For each dog, history, physical examination, complete blood count, biochemical profile, thoracic radiography, 6-lead standard electrocardiogram and trans-thoracic echocardiography were available. Comparisons were performed between groups of dogs belonging to different ACVIM stages and between dogs with and without PH. The median SDMA concentration was neither significantly different among groups of dogs in different disease stages (overall P = 0.010), nor among dogs with MMVD, nor between those with [14.5 μg/dl (10.5-18.8)] and without PH [13 μg/dl (9-17.2)] (P = 0.295). The concentration of SDMA did not differ between dogs when considering the combined effect of the ACVIM group and cardiac treatment (overall P = 0.486). Furthermore, no correlation was found between SDMA concentration and radiographic and echocardiographic parameters associated with increased MMVD severity. In conclusion, this study failed to demonstrate the presence of renal impairment in dogs with MMVD, and the increase in renal parameters in some dogs in the more advanced stage of MMVD could be attributed to pre-renal azotemia.