Mechanisms underlying the role of lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) in atherosclerotic development are not completely understood. We evaluated the relationship of Lp-PLA(2) with endothelial dysfunction, an early manifestation of atherosclerosis, in a cohort without known clinical cardiovascular disease. A total of 2809 participants in the Multi-Ethnic Study of Atherosclerosis underwent plasma Lp-PLA(2) mass and activity measurement and brachial artery flow-mediated vasodilation testing. In adjusted linear regression models, higher Lp-PLA(2) mass and activity levels were not associated with lower endothelial function (-0.04%, p = 0.51 and -0.09%, p = 0.10, respectively). Among individuals with subclinical atherosclerosis based on ankle-brachial index (ABI) or carotid intima-media thickness (IMT), Lp-PLA(2) mass and activity were not associated with lower endothelial function (-0.03%, p = 0.88 and -0.31%, p = 0.16 for ABI < 1.00; 0.01%, p = 0.94 and -0.15%, p = 0.20 for abnormal carotid IMT). In summary, Lp-PLA(2) is not associated with endothelial dysfunction, suggesting its role in atherosclerosis development is primarily related to other factors.