Atherosclerosis is a disease in which plaque builds up inside the arteries. During the early phase, endothelial nitric oxide nitrate (eNOS) decrease and nuclear factor kappa beta (NF-κβ) increase. Ambon banana peel contains XJP-1 and flavonoid which has been shown to inhibit the translocation of NF-κβ and increase eNOS expression in endothelial cells. This study was designed to investigate the effectiveness of Ambon Banana peel as atherosclerosis preventive agent through the inhibition of NF-κB and increasing eNOS expression in atherogenic rats. This study was post-test only control group design for 14 days using 30 male Wistar rats, divided into 5 groups: negative control with normal diet, positive control with atherogenic diet, and three control groups with atherogenic diet + 200 mg/kg body weight/ day, 400 mg/kg body weight/day, and 800 mg/kg body weight/day extract dosage. On the 15th day, the rats were terminated, taken its abdominal aortic and analyzed using immunohistochemical method. Statistical analysis showed that the extract significantly decreases NF-κβ (p=0.00) and increase eNOS (p=0.00). Pearson correlation proves that higher dose can lower NF-κ NF-κβ activity and increase eNOS expression (p<0.05). Linear regression shows that the extract can 82.1% lower NF-κβ activity and increase eNOS expression for 95.2%. In conclusion, the extract of Ambon banana peel has been proven to be effective in preventing atherosclerosis.