Abstract This study was conducted to determine the protective effects of peptide extract with 3 kDa from beef myofibrillar protein by Alkaline-AK enzyme (AK3K) against oxidative damage in SH-SY5Y cells and in the brains of spontaneously hypertensive rats (SHRs). All data were presented as the mean ± standard deviation of three replicates. All statistical analyses were performed using the one-way analysis of variance procedure in SPSS 20.0. A Tukey's multiple comparison test was used to determine significant differences between mean values, and evaluations were based on a significance level of P < 0.05. The AK3K did not affect cytotoxicity in SH-SY5Ycells (Figure 1). The resulting AK3K decrease the apoptotic ratio by 33.25%, DNA fragmentation, and reactive oxygen species (ROS) generation in oxidative damage induced SH-SY5Y cells (Figure 2). The SHR brains treated with the AK3K (400 mg/kg body weight, AK3K400) showed a significant decrease in malondialdehyde and ROS contents at 0.33 and 23.92 μM, respectively (P < 0.05) compared with the control. The SOD activity for AK3K was 61.26%, around 20% greater than the control. Therefore, this study suggest that the peptide extract from beef have neuroprotective effects such as inhibition of apoptotic factors and increment of antioxidant enzyme against oxidative stress and could possibly be used for neuronal hypertension therapy.