核桃油不皂化物对D-半乳糖诱导HepG2细胞的作用机制

Mechanism of Unsaponifiable Matter from Walnut Oil on D-Galactose-Induced HepG2 Cells

摘要:

衰老是一个不可避免的生物现象,包含多种生理变化。氧化应激是加剧衰老症状的主要原因之一,抵抗氧化应激是减轻衰老症状的有效手段。核桃油不皂化物是一种高活性成分,具有重要的开发和应用价值。采用D-半乳糖体外诱导HepG2细胞衰老模型,探讨核桃油不皂化物对HepG2细胞的作用,以细胞活力、细胞氧化应激水平、细胞内炎症因子的含量、细胞内肝功能水平评价不皂化物在D-半乳糖损伤模型上的作用,并进一步采用蛋白免疫印迹法探讨不皂化物对D-半乳糖损伤HepG2细胞的作用机制。结果表明:不皂化物(50、75、100μg/mL)处理显著改善了D-半乳糖损伤的HepG2细胞活力的降低,显著提高了超氧化物歧化酶(SOD)、谷胱甘肽过氧化酶(GPx)和过氧化氢酶(CAT)活性,降低了脂质氧化产物丙二醛(MDA)、炎症因子(TNF-α、IL-1β、IL-6)、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和碱性磷酸酶(AKP)的水平。此外,不皂化物促进了细胞质和细胞核中核因子E2相关因子2(Nrf2)的积累,激活了下游靶蛋白血红素氧合酶1(HO-1)和醌NADH脱氢酶1(NQO1)的表达。核桃油不皂化物延缓D-半乳糖诱导HepG2细胞的衰老与其激活Nrf2/Keap1信号通路有关,研究结果旨在为核桃油功能产品的开发提供理论参考。

Abstract:

Aging is an inevitable biological phenomenon that encompasses numerous physiological changes. Oxidative stress is one of the main reasons for exacerbating the symptoms of aging, and resistance to oxidative stress is an effective means of reducing the symptoms of aging. The unsaponifiable matter (USM) in walnut oil is a kind of highly active ingredient, which has important development and application value. The effect of USM from walnut oil on HepG2 cells was investigated using an in vitro D-galactose-induced senescence model of HepG2 cell and cell viability, cellular oxidative stress levels, intracellular inflammatory factor content, and intracellular liver function levels were used as evaluation indexes. The impact of USM on D-galactose-induced damage in HepG2 cells was examined using Western Blot analysis. The findings indicated that pre-treatment with USM at concentrations of 50,75, 100μg/mL effectively mitigated the reduced viability of D-galactose-damaged HepG2 cells, while also enhanced the enzymatic activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). The concentrations of lipid oxidation products, including malondialdehyde (MDA), inflammatory cytokines (TNF-α, IL-1β, IL-6), alanine and aspartate aminotransferase (ALT, AST), and alkaline phosphatase (AKP), exhibited a decrease. Furthermore, USM were found to enhance the translocation of nuclear factor E2-related factor 2 (Nrf2) into the cytoplasm and nucleus, leading to the activation of downstream target proteins such as heme oxygenase-1 (HO-1) and quinone NADH dehydrogenase 1 (NQO1). The USM of walnut oil have been shown to inhibit the senescence induced by D-galactose in HepG2 cells through activation of the Nrf2/Keap1 signaling pathway. The results aimed to provide theoretical references for the development of functional products from walnut oil.