Moon Ha-Rin, Yun Jung-Mi
Department of Food and Nutrition, Chonnam National University, Gwangju 61186, Korea.
Nutr Res Pract. 2023 Oct;17(5):899-916. doi: 10.4162/nrp.2023.17.5.899. Epub 2023 Jul 19.
BACKGROUND/OBJECTIVES: Oxidative stress is a fundamental neurodegenerative disease trigger that damages and decimates nerve cells. Neurodegenerative diseases are chronic central nervous system disorders that progress and result from neuronal degradation and loss. Recent studies have extensively focused on neurodegenerative disease treatment and prevention using dietary compounds. Heseperetin is an aglycone hesperidin form with various physiological activities, such as anti-inflammation, antioxidant, and antitumor. However, few studies have considered hesperetin's neuroprotective effects and mechanisms; thus, our study investigated this in hydrogen peroxide (HO)-treated SH-SY5Y cells.
MATERIALS/METHODS: SH-SY5Y cells were treated with HO (400 µM) in hesperetin absence or presence (10-40 µM) for 24 h. Three-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays detected cell viability, and 4',6-diamidino-2-phenylindole staining allowed us to observe nuclear morphology changes such as chromatin condensation and apoptotic nuclei. Reactive oxygen species (ROS) detection assays measured intracellular ROS production; Griess reaction assays assessed nitric oxide (NO) production. Western blotting and quantitative polymerase chain reactions quantified corresponding mRNA and proteins.
Subsequent experiments utilized various non-toxic hesperetin concentrations, establishing that hesperetin notably decreased intracellular ROS and NO production in HO-treated SH-SY5Y cells ( < 0.05). Furthermore, hesperetin inhibited HO-induced inflammation-related gene expression, including interluekin-6, tumor necrosis factor-α, and nuclear factor kappa B (NF-κB) p65 activation. In addition, hesperetin inhibited NF-κB translocation into HO-treated SH-SY5Y cell nuclei and suppressed mitogen-activated protein kinase protein expression, an essential apoptotic cell death regulator. Various apoptosis hallmarks, including shrinkage and nuclear condensation in HO-treated cells, were suppressed dose-dependently. Additionally, hesperetin treatment down-regulated Bax/Bcl-2 expression ratios and activated AMP-activated protein kinase-mammalian target of rapamycin autophagy pathways.
These results substantiate that hesperetin activates autophagy and inhibits apoptosis and inflammation. Hesperetin is a potentially potent dietary agent that reduces neurodegenerative disease onset, progression, and prevention.
背景/目的：氧化应激是引发神经退行性疾病的一个基本因素，它会损害并大量杀死神经细胞。神经退行性疾病是慢性中枢神经系统疾病，由神经元退化和丧失所致且会不断发展。最近的研究广泛聚焦于使用膳食化合物治疗和预防神经退行性疾病。橙皮素是一种具有多种生理活性的橙皮苷苷元形式，如抗炎、抗氧化和抗肿瘤活性。然而，很少有研究探讨橙皮素的神经保护作用及其机制；因此，我们的研究在过氧化氢（H₂O₂）处理的SH-SY5Y细胞中对此进行了研究。
材料/方法：在不存在或存在橙皮素（10 - 40 μM）的情况下，用H₂O₂（400 μM）处理SH-SY5Y细胞24小时。噻唑蓝检测细胞活力，4',6-二脒基-2-苯基吲哚染色使我们能够观察核形态变化，如染色质浓缩和凋亡细胞核。活性氧（ROS）检测测定细胞内ROS产生；Griess反应测定评估一氧化氮（NO）产生。蛋白质免疫印迹法和定量聚合酶链反应对相应的mRNA和蛋白质进行定量。
随后的实验使用了各种无毒的橙皮素浓度，证实橙皮素显著降低了H₂O₂处理的SH-SY5Y细胞内的ROS和NO产生（P < 0.05）。此外，橙皮素抑制H₂O₂诱导的炎症相关基因表达，包括白细胞介素-6、肿瘤坏死因子-α和核因子κB（NF-κB）p65的激活。此外，橙皮素抑制NF-κB转位到H₂O₂处理的SH-SY5Y细胞核中，并抑制丝裂原活化蛋白激酶蛋白表达，这是一种重要的凋亡细胞死亡调节因子。各种凋亡特征，包括H₂O₂处理细胞中的细胞收缩和核浓缩，均呈剂量依赖性受到抑制。此外，橙皮素处理下调了Bax/Bcl-2表达比率，并激活了AMP活化蛋白激酶-雷帕霉素哺乳动物靶标自噬途径。
这些结果证实橙皮素激活自噬并抑制凋亡和炎症。橙皮素是一种潜在的有效膳食剂，可减少神经退行性疾病的发生、发展并起到预防作用。
