Protective effect of Buddha’s Temple extract against tert-butyl hydroperoxide stimulation-induced oxidative stress in DF-1 cells

Abstract

Objective: To determine the protective efficacy of Buddha's Temple (BT) extract against tertbutyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus DF-1 fibroblast cells and its effects on the cell lipid metabolism. Methods:In this experimental study, Gallus gallus DF-1 fibroblast cells were pretreated with BT 10 -7 for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). WST8 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. Results:The WST8 assay results showed that the BT pretreatment of Gallus gallus DF-1 fibroblast cell increased their cell survival rate by 1.08 ± 0.04%, decreased the 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 TNF α, TICAM1, and GRP78 were regulated, which contributed to cell stabilization. TLC and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as TAG (triacylglycerol) and FFA (free fatty acids). Conclusion:In this study, we confirmed that the examined BT extract exerted selective protective effects on Gallus gallus DF-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.