Park Eun Hye, Kim Sung-Jo
Division of Cosmetics and Biotechnology, College of Life and Health Sciences, Hoseo University, Baebang, Asan 31499, Korea.
Anim Biosci. 2023 Jul;36(7):1120-1129. doi: 10.5713/ab.23.0014. Epub 2023 Feb 27.
This study aimed to determine the protective efficacy of Buddha's Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus chicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism.
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). 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.
The WST-8 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 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 factor α (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.
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.
本研究旨在确定佛寺庙提取物（BT）对叔丁基过氧化氢（t-BHP）诱导的原鸡鸡胚成纤维细胞系（DF-1）氧化应激的保护作用及其对细胞脂质代谢的影响。
在本实验研究中，将原鸡DF-1成纤维细胞用10⁻⁷的BT预处理24小时，然后用100μM的t-BHP处理6小时。进行水溶性四氮唑盐-8（WST-8）检测，并计算生长曲线。通过定量聚合酶链反应（qPCR）确认BT提取物引起的细胞内基因表达变化。进行流式细胞术、油红O染色实验和薄层色谱法检测细胞内代谢机制变化。
WST-8检测结果表明，BT预处理原鸡DF-1成纤维细胞可使其细胞存活率提高(1.08%\pm0.04%)，即使在暴露于氧化剂后，活性氧（ROS）水平降低(0.93%\pm0.12%)，线粒体活性稳定(1.37%\pm0.36%)。此外，qPCR结果证实肿瘤坏死因子α（TNFα）、含TIR结构域的衔接蛋白诱导IFN-β（TICAM1）和葡萄糖调节蛋白78（GRP78）的基因表达水平受到调节，这有助于细胞稳定。薄层色谱法和油红O分析显示三酰甘油和游离脂肪酸等脂质代谢产物的含量明显降低。
在本研究中，我们证实所检测的BT提取物对原鸡DF-1成纤维细胞免受强氧化诱导剂t-BHP引起的细胞损伤具有选择性保护作用。此外，我们确定该提取物显著降低了氧化应激导致的细胞内ROS积累，这有助于提高家禽产量和增加收入。
