Zhongguo youzhi (Sep 2024)
紫苏多糖体外抗氧化活性及对肝细胞 氧化损伤的保护作用In vitro antioxidant activity of polysaccharide from perilla seed meal and its protective effects against oxidative stress-induced damage of hepatocytes
Abstract
旨在为紫苏饼粕的高值化利用提供参考，以采用水提醇沉法从紫苏粕中提取的紫苏多糖（PMP）为原料，检测其对羟自由基（·OH）、超氧阴离子自由基（O-2·）、1，1-二苯基-2-三硝基苯肼自由基（DPPH·）和亚硝酸盐（NO-2）的清除能力，考察其体外抗氧化活性；同时，使用H2O2建立LO2细胞氧化损伤模型，在细胞水平上探讨PMP的抗氧化能力，并进一步研究PMP对LO2肝细胞氧化应激损伤的保护作用机制。结果表明：PMP对·OH、O-2·、DPPH·、NO-2均具有一定的清除能力，其EC50分别为964.59、6 376.84、333.55、275.24 μg/mL；LO2细胞氧化损伤模型的构建条件为H2O2浓度800 μmol/L处理时间24 h；PMP可显著提高H2O2诱导损伤的LO2细胞活力，当PMP质量浓度为1 000 μg/mL时，细胞活力可提高到92.46%；在保护LO2细胞氧化损伤机制方面，PMP显著降低H2O2诱导的细胞内活性氧（ROS）生成水平，提升线粒体膜电位，提高细胞内谷胱甘肽（GSH）含量以及超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性，减少丙二醛(MDA）的含量，并通过显著上调凋亡蛋白Bcl-2的表达和下调Bax的表达改善氧化应激损伤引起的细胞凋亡，提高细胞的增殖活性。综上，PMP表现出一定的自由基清除活性以及对H2O2氧化损伤的LO2细胞的保护作用。To provide reference for the high-value utilization of perilla seed meal, perilla polysaccharide (PMP) prepared from perilla seed meal by water solution and alcohol precipitation was used as material, and its in vitro antioxidant activity was analyzed by detecting the scavenging rate of hydroxyl free radical (·OH), superoxide anion radical(O-2·), 1,1-diphenyl-2- picrylhydrazyl radical (DPPH·), nitrite (NO-2). At the same time, H2O2 was used to establish LO2 hepatocyte damage model, and the antioxidant ability of PMP was investigated in cellular level. The protective mechanism of PMP on LO2 hepatocytes oxidative stress damage was further studied. The results showed that PMP had certain scavenging ability to ·OH, O-2·, DPPH· and NO-2, and their EC50 values were 964.59，6 376.84，333.55，275.24 μg/mL, respectively. A concentration of 800 μmol/L H2O2 treatment for 24 h was selected as the construction condition for the oxidative damage model of LO2 cells. PMP could significantly increase the viability of LO2 cells with H2O2-induced damage, and the cell viability increased to 92.46% when the PMP mass concentration was 1 000 μg/mL. In terms of the mechanism of protecting LO2 cells from oxidative damage, PMP significantly reduced the level of intracellular reactive oxygen species (ROS) induced by H2O2, increased mitochondrial membrane potential, increased the content of glutathione (GSH) and the activities of superoxide dismutase (SOD) and catalase (CAT) in cells, reduced the level of malondialdehyde (MDA), and improved the apoptosis caused by oxidative stress damage by significantly up-regulating the expression of apoptotic protein Bcl-2 and down-regulating the expression of Bax. In summary, PMP exhibit certain free radical scavenging activity and protective effects on LO2 cells damaged by H2O2.
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