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To examine the associations of resistance exercise, independent of and combined with aerobic exercise, with the risk of development of hypercholesterolemia in men.This study used data from the Aerobics Center Longitudinal Study, which is a cohort examining the associations of clinical and lifestyle factors with the development of chronic diseases and mortality. Participants received extensive preventive medical examinations at the Cooper Clinic in Dallas, Texas, between January 1, 1987, and December 31, 2006. A total of 7317 men aged 18 to 83 years (mean age, 46 years) without hypercholesterolemia at baseline were included. Frequency (times per week) and total amount (min/wk) of resistance and aerobic exercise were determined by self-report. Hypercholesterolemia was defined as a total cholesterol level of 240 mg/dL or higher or physician diagnosis.During a median (interquartile range) follow-up of 4 (2 to 7) years, hypercholesterolemia developed in 1430 of the 7317 men (20%). Individuals meeting the resistance exercise guidelines (≥2 d/wk) had a 13% lower risk of development of hypercholesterolemia (hazard ratio [HR], 0.87; 95% CI, 0.76-0.99; P=.04) after adjustment for general characteristics, lifestyle factors, and aerobic exercise. In addition, less than 1 h/wk and 2 sessions per week of resistance exercise were associated with 32% and 31% lower risks of hypercholesterolemia (HR, 0.68; 95% CI, 0.54-0.86; P=.001; and HR, 0.69; 95% CI, 0.54-0.88; P=.003), respectively, compared with no resistance exercise. Higher levels of resistance exercise did not provide benefits. Meeting both resistance and aerobic exercise guidelines (≥500 metabolic equivalent task min/wk) lowered the risk of development of hypercholesterolemia by 21% (HR, 0.79; 95% CI, 0.68-0.91; P=.002). compared with meeting none of the guidelines.Compared with no resistance exercise, less than 1 h/wk of resistance exercise, independent of aerobic exercise, is associated with a significantly lower risk of development of hypercholesterolemia in men (P=.001). However, the lowest risk of hypercholesterolemia was found at 58 min/wk of resistance exercise. This finding suggests that resistance exercise should be encouraged to prevent hypercholesterolemia in men. However, future studies with a more rigorous analysis including major potential confounders (eg, diet, medications) are warranted.