神经酸对Hydrogen peroxide（H2O2）诱导PC12细胞氧化损伤的保护作用

摘要/Abstract

摘要： 摘要：神经酸（Nervonic acid，NA）是一种超长链单不饱和脂肪酸，在神经细胞发育中起着关键作用，具有潜在的利用价值。本研究以PC12细胞（大鼠嗜铬细胞瘤细胞系）作为研究对象，采用过氧化氢（hydrogen peroxide，H2O2）为诱导剂，确定H2O2最佳处理浓度和时间，建立细胞氧化损伤模型。同时，研究NA对PC12细胞存活率、超氧化物歧化酶（Superoxide dismutase，SOD）、谷胱甘肽过氧化物酶 (Glutathione peroxidase，GSH–PX)活性，乳酸脱氢酶（lactate dehydrogenase，LDH）和丙二醛（Malondialdehyde，MDA）含量的影响，并对细胞凋亡和细胞内活性氧(reactive oxygen species，ROS)水平进行评估。采用蛋白印迹（Western blotting，WB）和实时荧光定量技术（quantitative real time PCR, RT-qPCR）检测BCL2-Associated X的蛋白质（Bax）、B淋巴细胞瘤-2基因（B-cell lymphoma-2，Bcl-2）、半胱氨酸蛋白酶-3（Caspase-3）、核因子E2相关因子2（Nuclear Factor erythroid 2-Related Factor 2，Nrf2）、kelch 样ECH 关联蛋白1（kelch-like ECH-associated protein 1, Keap1）和血红素加氧酶-1（heme oxygenase 1，HO-1）等细胞凋亡和氧化应激有关蛋白和基因的表达水平。结果表明，使用终浓度为200μΜ的 H2O2处理PC12细胞24h后，对细胞的抑制率为60.12%；细胞毒性试验表明，终浓度为10μΜ以下的NA对细胞无毒副作用；NA能够显著降低H2O2诱导损伤细胞中的LDH和MDA含量、ROS的过度产生，并增强SOD和GSH-PX抗氧化酶活性，显著上调Bcl-2、Nrf2和HO-1蛋白和基因表达水平，下调Caspase-3、Bax和Keap1蛋白和基因表达水平。综上所述， NA对H2O2诱导的PC12细胞氧化损伤具有保护作用，且与激活Keap1/Nrf2/HO-1信号通路有关。

关键词: 关键词：神经酸, 氧化损伤, PC12细胞(大鼠肾上腺髓质嗜铬细胞瘤), 信号通路

Abstract: Abstract: Nervonic acid (NA) is a kind of ultra long chain single unsaturated fat fatty acid, which plays a key role in the development of nerve cells and has potential utilization value. This study focuses on PC12 cells (rat pheochromocytoma cell line), using hydrogen peroxide (H2O2) as an inducer to determine the optimal treatment concentration and time of H2O2, and establish a cell oxidative damage model. At the same time, the effects of NA on PC12 cell survival rate, superoxide dismutase (SOD), glutathione peroxidase (GSH–PX) activity, lactate dehydrogenase (LDH) and malondialdehyde (MDA) content were studied, and the levels of apoptosis and reactive oxygen species (ROS) in cells were evaluated. Western blotting (WB) and real-time fluorescence quantitative PCR (RT- qPCR) were used to detect the expression levels of proteins and genes related to cell apoptosis and oxidative stress, such as BCL2 Associated X (Bax), B-cell lymphoma-2 (Bcl-2), Caspase-3, Nuclear Factor E2 Related Factor 2 (Nrf2), kelch like ECH associated protein 1 (Keap1), and heme oxygenase-1 (HO-1). The results indicate that the final concentration used is 200 μΜ After treating PC12 cells with H2O2 for 24 hours, the cell survival rate was 60.12%; The cytotoxicity test showed that the final concentration was 10μΜ the following NA has no toxic side effects on cells; NA can significantly reduce the content of LDH and MDA, excessive production of ROS, and enhance the activity of SOD and GSH-PX antioxidant enzymes in H2O2 induced injury cells. It significantly upregulates the expression levels of Bcl-2, Nrf2, and HO-1 proteins and genes, Downregulate the protein and gene expression levels of Caspase-3, Bax and Keap1. In summary, NA has a protective effect on H2O2 induced oxidative damage in PC12 cells and is associated with the activation of the Nrf2/ Keap1/HO-1 signaling pathway.

Key words: Key words: Nervonic acid, Oxidative damage, PC12 cells (Pheochromocytoma cells), Signaling pathway

中图分类号:

q291

玛依乐·艾海提刘垚杰李建科. 神经酸对Hydrogen peroxide（H2O2）诱导PC12细胞氧化损伤的保护作用[J]. 食品科学.

玛依乐艾海提 Jianke Li. Protective effect of nervonic acid on oxidative damage of PC12 cells induced by hydrogen peroxide (H2O2)[J]. FOOD SCIENCE.

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