In this study, the nootropic, antifatigue and antioxidant effects of two commercial milk fat globule membranes (MFGM) in BALB/c male mice were evaluated and the possible mechanisms were elucidated. Normal saline was used a control. The experimental results showed that MFGM could improve learning and cognitive ability spatial memory ability in mice, extend exhaustive swimming time, significantly increase glycogen levels, and greatly improve fatigue- and oxidative stress-related indicators, indicating its nootropic, antifatigue and antioxidant effects. However, the effects of the two MFGMs, which are produced by different methods and have different compositions, were different.