Sleep is an important physiological behavior in humans that is associated with the occurrence and development of various diseases. However, the association of sleep duration with health-related outcomes, including obesity-related factors, musculoskeletal diseases, and mortality because of different causes, has not been systematically reported. This study aims to systematically investigate the effect of sleep duration on health-related outcomes. Overall, 54,664 participants with sleep information from 8 survey cycles of the National Health and Nutrition Examination Survey (2005-2020) were included in the analysis. Health-related outcomes comprised obesity-related outcomes (ie, BMI, obesity, waist circumference, and abdominal obesity), metabolism-related outcomes (ie, uric acid, hyperuricemia, and bone mineral density [BMD]), musculoskeletal diseases (ie, osteoarthritis [OA] and rheumatoid arthritis [RA]), and mortality because of different causes. The baseline information of participants including age, sex, race, educational level, marital status, total energy intake, physical activity, alcohol consumption, smoking, hypertension, and diabetes was also collected as covariates. Information about the metabolism index, disease status, and covariates was acquired from the laboratory, examination, and questionnaire data. Survival information, including survival status, duration, and cause of death, was obtained from the National Death Index records. Quantile regression models and Cox regression models were used for association analysis between sleep duration and health-related outcomes. In addition, the threshold effect analysis, along with smooth curve fitting method, was applied for the nonlinear association analysis. Participants were divided into 4 groups with different sleep durations. The 4 groups showed significant differences in terms of baseline data (P<.001). The quantile regression analysis indicated that participants with increased sleep duration showed decreased BMI (β=-.176, 95% CI -.220 to -.133; P<.001), obesity (odds ratio [OR] 0.964, 95% CI 0.950-0.977; P<.001), waist circumference (β=-.219, 95% CI -.320 to -.117; P<.001), abdominal obesity (OR 0.975, 95% CI 0.960-0.990; P<.001), OA (OR 0.965, 95% CI 0.942-0.990; P=.005), and RA (OR 0.940, 95% CI 0.912-0.968; P<.001). Participants with increased sleep duration also showed increased BMD (β=.002, 95% CI .001-.003; P=.005), as compared with participants who slept <5.5 hours. A significant saturation effect of sleep duration on obesity, abdominal obesity, and hyperuricemia was detected through smooth curve fitting and threshold effect analysis (sleep duration>inflection point). In addition, a significant threshold effect of sleep duration on BMD (P<.001); OA (P<.001); RA (P<.001); and all-cause (P<.001), cardiovascular disease-cause (P<.001), cancer-cause (P=.005), and diabetes-cause mortality (P<.001) was found. The inflection point was between 6.5 hours and 9 hours. The double-edged sword effect of sleep duration on obesity-related outcomes, embolism-related diseases, musculoskeletal diseases, and mortality because of different causes was detected in this study. These findings provided epidemiological evidence that proper sleep duration may be an important factor in the prevention of multisystem diseases.