Background: Natriuretic peptides (NPs), including brain natriuretic peptide (BNP), are neurohormones involved in the regulation of water-sodium balance and the maintenance of cardiovascular homeostasis. A higher concentration of NPs is observed in females, but the mechanism behind this difference has not been fully elucidated. Methods: Randomly chosen 255 volunteers from the general population were examined. Overall, 196 people without severe cardiovascular disease were included (mean age 48 years, 35.7% male). A comprehensive assessment was performed, including anthropometric measurements, N-terminal pro-brain natriuretic peptide (NT-proBNP), total testosterone (TT) and sex hormone-binding globulin (SHBG) concentration, transthoracic echocardiography (ECHO), and body composition analysis by direct dual-energy X-ray absorptiometry (DEXA). The univariate analysis adjusted by the known affecting factors determined which measurements were independently associated with NT-proBNP concentration. Results: NT-proBNP concentration was positively associated with gynoid fat mass, gynoid/total fat (G/TF) mass index, SHBG and negatively with android/gynoid (A/G) fat mass index, TT and calculated free testosterone (CFT) concentrations. Furthermore, body composition parameters remained independently associated with NT-proBNP levels even after adjusting for CFT and SHBG. Conclusion: In the population without severe cardiovascular disease, the NT-proBNP concentration is independently associated with lower availability of testosterone and higher gynoid fat distribution, which may explain higher NPs levels in females.