ObjectiveThis study aimed to determine the protective efficacy of Buddha’s Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress inGallus galluschicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism.MethodsIn this experimental study,Gallus gallusDF-1 fibroblast cells were pretreated with BT 10−7for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). Water-soluble tetrazolium salt-8 (WST-8) assays were performed, and the growth curve was computed. The intracellular gene expression changes caused by BT extract were confirmed through quantitative polymerase chain reaction (qPCR). Flow cytometry, oil red O staining experiment, and thin-layer chromatography were performed for the detection of intracellular metabolic mechanism changes.ResultsThe WST-8 assay results showed that the BT pretreatment ofGallus gallusDF-1 fibroblast cell increased their cell survival rate by 1.08%±0.04%, decreased the reactive oxygen species (ROS) level by 0.93%±0.12% even after exposure to oxidants, and stabilized mitochondrial activity by 1.37%±0.36%. In addition, qPCR results confirmed that the gene expression levels of tumor necrosis factorα(TNFα), TIR domain-containing adapter inducing IFN-beta (TICAM1), and glucose-regulated protein 78 (GRP78) were regulated, which contributed to cell stabilization. Thin-layer chromatography and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as triacylglycerol and free fatty acids.ConclusionIn this study, we confirmed that the examined BT extract exerted selective protective effects onGallus gallusDF-1 fibroblast cells against cell damage caused by t-BHP, which is a strong oxidative inducer. Furthermore, we established that this extract significantly reduced the intracellular ROS accumulation due to oxidative stress, which contributes to an increase in poultry production and higher incomes.