食品与发酵工业 >

2024 , Vol. 50 >Issue 8: 105 - 113

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035027

研究报告

桑葚花色苷对PC12细胞氧化损伤的保护作用

  • 朱晓晗 ,
  • 铁芳芳 ,
  • 欧阳健 ,
  • 王洪伦 ,
  • 殷军港
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  • 1(烟台大学 生命科学学院,山东 烟台,264005)
    2(中国科学院藏药研究重点实验室,西北高原生物研究所,青海 西宁,810008)
    3(青海省藏药研究重点实验室,青海 西宁,810008)
第一作者:硕士研究生(殷军港教授为通信作者,E-mail:ytuyinjungang@126.com)

收稿日期: 2023-02-09

  修回日期: 2023-04-04

  网络出版日期: 2024-06-11

基金资助

青海省自然科学基金面上项目(2020-ZJ-905)

收起

Protective effects of mulberry anthocyanin on oxidative stress in PC12 cells

  • ZHU Xiaohan ,
  • TIE Fangfang ,
  • OUYANG Jian ,
  • WANG Honglun ,
  • YIN Jungang
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  • 1(College of Life Sciences, Yantai University, Yantai 264005, China)
    2(Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, China)
    3(Qinghai Provincial Key Laboratory of Tibetan Medicine Research, Xining 810008, China)

Received date: 2023-02-09

  Revised date: 2023-04-04

  Online published: 2024-06-11

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摘要

通过建立大鼠肾上腺嗜铬细胞瘤细胞(PC12细胞)氧化应激损伤模型,探讨桑葚花青素中2个矢车菊素类化合物即矢车菊素-3-O-葡萄糖苷(cyanidin-3-O-glucoside,C3G)和矢车菊素-3-O-芸香糖苷(cyanidin-3-O-rutinoside,C3R)对过氧化氢(H2O2)诱导的细胞氧化损伤的保护作用及相关作用机制。采用噻唑蓝法检测细胞活力,DCFH-DA荧光探针检测活性氧(reactive oxygen species,ROS)的含量,试剂盒测定细胞中还原型谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)以及过氧化氢酶(catalase,CAT)的水平。Western Blot检测抗氧化以及细胞凋亡蛋白的表达水平。结果表明,650 μmol/L H2O2处理PC12细胞12 h后,细胞存活率显著降低(P<0.01),50、100 μmol/L的C3G、C3R能显著升高细胞活力(P<0.05)及GSH、SOD和CAT等抗氧化酶的活性(P<0.05),降低细胞内ROS以及MDA的含量。此外50、100 μmol/L的C3G、C3R处理后显著抑制PC12细胞内c-Jun氨基末端激酶和细胞外调节蛋白激酶蛋白磷酸化水平,抑制半胱氨酸天冬氨酸蛋白酶-3以及Bcl-2相关X蛋白等凋亡蛋白的表达(P<0.05),促进核因子E-2-相关因子和血红素加氧酶1抗氧化蛋白的表达水平。综上,C3G、C3R对H2O2所致的PC12细胞氧化损伤具有保护作用,可能与抑制细胞凋亡通路相关蛋白的表达,提高胞内抗氧化酶活性,清除胞内过量ROS,降低细胞凋亡有关。

关键词： 桑葚花色苷; 矢车菊素-3-O-葡萄糖苷; 矢车菊素-3-O-芸香糖苷; PC12细胞; 氧化应激; 细胞凋亡

本文引用格式

朱晓晗 , 铁芳芳 , 欧阳健 , 王洪伦 , 殷军港 . 桑葚花色苷对PC12细胞氧化损伤的保护作用[J]. 食品与发酵工业, 2024 , 50(8) : 105 -113 . DOI: 10.13995/j.cnki.11-1802/ts.035027

Abstract

The protective function and related mechanism against H2O2-induced cell injury of cyanidin compounds cyanidin-3-O-glucoside (C3G) and cyanidin-3-O-rutinoside (C3R) was investigated by establishing the model of an oxidative stress injury in PC12 cells.MTT assay and DCFH-DA fluorescent probe were employed to detect cell viability and reactive oxygen species (ROS) content, respectively.The test kits were applied to detect reduced glutathione (GSH), malondialdehyde (MDA) content, superoxide dismutase (SOD), and catalase (CAT) activity.Antioxidant and apoptotic protein expression level was detected by Western blot.Results showed that the cell survival rate was significantly decreased treated with 650 μmol/L H2O2 for 12 hours (P<0.01), 50 μmol/L and 100 μmol/L C3G and C3R could improve the survival rate of PC12 cells, inhibit the production of ROS, decrease the content of MDA, and enhance the activities of GSH, SOD, and CAT in PC12 cells compared with H2O2 treatment group (P<0.05).Western blot results showed that 50 μmol/L and 100 μmol/L C3G and C3R could significantly inhibit the phosphorylation levels of c-Jun N-terminal kinase (JNK) and extracellular regulated protein kinases (ERK1/2) proteins as well as the expression of Caspase-3 and Bcl-2-associated X protein (Bax), and promote the protein levels of nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) (P<0.05).In summary, C3G and C3R have protective effects on H2O2-induced oxidative damage in PC12 cells which may be related to inhibiting the expression of apoptosis-related proteins, improving the activity of intracellular antioxidant enzymes, clearing excess ROS in cells, and reducing the rate of apoptosis.

Key words： mulberry anthocyanin; cyanidin-3-O-glucoside; cyanidin-3-O-rutinoside; PC12 cells; oxidative stress; apoptosis

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