Xu Yifan, Wang Yawen, Yang Min, Yuan Pengxiang, Xu Weikang, Jiang Tong, Huang Jian
National Institute for Nutrition and Health, Chinese Center for Disease Control and Prevention, Beijing 100050, China.
Key Laboratory of Public Nutrition and Health, National Health Commission of the People's Republic of China, Beijing 100050, China.
Nutrients. 2025 Jul 2;17(13):2204. doi: 10.3390/nu17132204.
Under oxidative stress conditions, the increased levels of reactive oxygen species (ROS) within cells disrupt the intracellular homeostasis. Tartary buckwheat peptides exert their effects by scavenging oxidative free radicals, such as superoxide anion and hydrogen peroxide, thereby reducing oxidative damage within cells. Meanwhile, these peptides safeguard mitochondria by maintaining the mitochondrial membrane potential, decreasing the production of mitochondrial oxygen free radicals, and regulating mitochondrial biogenesis and autophagy to preserve mitochondrial homeostasis. Through these mechanisms, Tartary buckwheat peptides restore the intracellular redox balance, sustain cellular energy metabolism and biosynthesis, and ensure normal cellular physiological functions, which is of great significance for cell survival and adaptation under oxidative stress conditions. In this experiment, a classical cellular oxidative stress model was established. Indicators related to antioxidant capacity and mitochondrial membrane potential changes, as well as pathways associated with oxidative stress, were selected for detection. The aim was to elucidate the effects of Tartary buckwheat oligopeptides on the metabolism of cells in response to oxidative stress. In this study, we established an oxidative damage model of mouse skeletal muscle myoblast (SOL8) cells using hydrogen peroxide (HO), investigated the pre-protective effects of Tartary buckwheat oligopeptides on HO-induced oxidative stress damage in SOL8 cells at the cellular level, and explored the possible mechanisms. The CCK-8 method is a colorimetric assay based on WST-8-[2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodiumsalt], which is used to detect cell proliferation and cytotoxicity. The value of CCK-8 showed that, when the cells were exposed to 0.01 mmol/L HO for 1 h and 10 mg/mL Tartary buckwheat oligopeptides intervention for 48 h, these were the optimal conditions. Compared with the HO group, the intervention group (KB/HO group) showed that the production of ROS was significantly reduced ( < 0.001), the malondialdehyde (MDA) content was significantly decreased ( < 0.05), and the activity of catalase (CAT) was significantly increased ( < 0.01); the mitochondrial membrane potential in the KB/HO group tended to return to the level of the control group, and they all showed dose-dependent effects. Compared with the HO group, the mRNA expression of KEAP1 in the KB/HO group decreased, while the mRNA expression of NRF2α, HO-1, nrf1, PGC-1, P62, and PINK increased. : Therefore, Tartary buckwheat oligopeptides have a significant pre-protective effect on HO-induced SOL8 cells, possibly by enhancing the activity of superoxide dismutase, reducing ROS attack, balancing mitochondrial membrane potential, and maintaining intracellular homeostasis.
在氧化应激条件下,细胞内活性氧(ROS)水平升高会破坏细胞内稳态。苦荞肽通过清除超氧阴离子和过氧化氢等氧化自由基发挥作用,从而减少细胞内的氧化损伤。同时,这些肽通过维持线粒体膜电位、减少线粒体氧自由基的产生以及调节线粒体生物发生和自噬来保护线粒体,以维持线粒体稳态。通过这些机制,苦荞肽恢复细胞内氧化还原平衡,维持细胞能量代谢和生物合成,并确保细胞正常的生理功能,这对于氧化应激条件下细胞的存活和适应具有重要意义。在本实验中,建立了经典的细胞氧化应激模型。选择与抗氧化能力和线粒体膜电位变化相关的指标以及与氧化应激相关的信号通路进行检测。目的是阐明苦荞寡肽对氧化应激下细胞代谢的影响。在本研究中,我们用过氧化氢(H₂O₂)建立了小鼠骨骼肌成肌细胞(SOL8)的氧化损伤模型,在细胞水平上研究了苦荞寡肽对H₂O₂诱导的SOL8细胞氧化应激损伤的预保护作用,并探讨了可能的机制。CCK-8法是一种基于WST-8-[2-(2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺酸苯基)-2H-四唑鎓,单钠盐]的比色法,用于检测细胞增殖和细胞毒性。CCK-8值表明,当细胞暴露于0.01 mmol/L H₂O₂ 1小时并经10 mg/mL苦荞寡肽干预48小时时,这些是最佳条件。与H₂O₂组相比,干预组(KB/H₂O₂组)显示ROS产生显著减少(P<0.001),丙二醛(MDA)含量显著降低(P<0.05),过氧化氢酶(CAT)活性显著增加(P<0.01);KB/H₂O₂组的线粒体膜电位趋于恢复到对照组水平,且均呈现剂量依赖性效应。与H₂O₂组相比,KB/H₂O₂组中KEAP1的mRNA表达下降,而NRF2α、HO-1、nrf1、PGC-1、P62和PINK的mRNA表达增加。因此,苦荞寡肽对H₂O₂诱导的SOL8细胞具有显著的预保护作用,可能是通过增强超氧化物歧化酶的活性、减少ROS攻击、平衡线粒体膜电位以及维持细胞内稳态来实现的。