Purpose: DNA damage and repair responses are induced by metabolic diseases and environmental stress.The balance of DNA repair response and the antioxidant system play a role in modulating the entire body's health.This study uses a high-fat and high-calorie (HFC) drink to examine the new roles of a plant-based multivitamin/mineral supplement with phytonutrients (PMP) for regulating the antioxidant system and cellular DNA repair signaling in the body resulting from metabolic stress.Methods: In a double-blind, randomized, parallel-arm, and placebo-controlled trial, healthy adults received a capsule containing either a PMP supplement (n = 12) or a placebo control (n = 12) for 8 wk.Fasting blood samples were collected at 0, 1, and 3 h after consuming a HFC drink (900 kcal).The blood samples were analyzed for the following oxidative stress makers: areas under the curve reactive oxygen species (ROS) levels, plasma malondialdehyde (MDA), erythrocytes MDA, urinary MDA, oxidized low-d. lipoprotein, and the glutathione:oxidized glutathione ratio at the time points.We further examined the related protein levels of DNA repair signaling (pCHK1 (Serine 345), p-P53 (Serine 15), and ΓH2AX expression) in the plasma of subjects to evaluate the time-dependent effects of a HFC drink.Results: In a previous study, we showed that PMP supplementation for eight weeks reduces the ROS and endogenous DNA damage in human blood plasma.Results of the current study further show that PMP supplementation is significantly correlated with antioxidant defense.Compared to the placebo samples, the blood plasma obtained after PMP supplementation showed enhanced DNA damage response genes such as pCHK1(Serine 345) (a transducer of DNA response) and γH2AX (a hallmark of DNA damage) during the 8 wk trial on metabolic challenges.Conclusion: Our results indicate that PMP supplementation for 8 wk enhances the anti-oxidant system against oxidative stress and prevents DNA damage signaling in humans.