食品科学

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不同黄酮类单体对CHO细胞早期损伤作用的比较研究

刘 涛,孙 健,郭 辰,赵晓红   

  1. 1.首都师范大学生命科学学院,北京 100048;2.北京联合大学功能食品研究院,北京 100191
  • 出版日期:2014-06-15 发布日期:2014-07-03

Comparison of Early Damage of CHO Cells in the Presence of Different Flavonoid Monomers

LIU Tao, SUN Jian, GUO Chen, ZHAO Xiao-hong   

  1. 1.School of Life Science, Capital Normal University, Beijing 100048, China;
    2.Research Institute for Science and Technology of Functional Foods, Beijing Union University, Beijing 100191, China
  • Online:2014-06-15 Published:2014-07-03

摘要:

选用竹叶中4 种黄酮单体荭草苷、异荭草苷、绿原酸和阿魏酸,在各毒性剂量下作用于中国仓鼠卵巢(Chinese hamster ovary,CHO)细胞,通过检测其对细胞早期损伤指标——线粒体功能变化的影响,以评估其细胞损伤作用。CCK-8检测法测得4 种单体物质对CHO细胞的IC50值分别是:绿原酸2 726.11 μmol/L、阿魏酸1 680.28 μmol/L、荭草苷1 889.15 μmol/L、异荭草苷2 358.56 μmol/L。在各单体IC50、1/2 IC50值、1/4 IC50值、1/8IC50值剂量下,线粒体膜电位变化具有剂量依赖性,其中以阿魏酸降低幅度最明显,绿原酸次之,荭草苷和异荭草苷变化较小;三磷酸腺苷(adenosine triphosphate,ATP)生成量与作用剂量成负相关,阿魏酸致ATP生成量降低最多,荭草苷次之,绿原酸和异荭草苷影响较小;细胞色素C的释放量,以阿魏酸最大,荭草苷和异荭草苷次之,绿原酸的最小。研究表明4 种单体物质都对线粒体产生了一定的损伤,其中阿魏酸对线粒体影响最大,荭草苷和异荭草苷次之,绿原酸最小。

关键词: 细胞色素C, 线粒体膜电位, 三磷酸腺苷, 线粒体损伤, 绿原酸, 阿魏酸, 荭草苷, 异荭草苷

Abstract:

The cytotoxicity of four types of flavonoid monomers, namely chlorogenic acid, ferulic acid, orientin and
isoorientin on Chinese hamster ovary (CHO) cells was detected and compared by CCK-8 test and the IC50 values were
calculated by Exact Probability. The mitochondrial membrane potential, the production of ATP and the release of cytochrome
C were used as the parameters to indicate early cellular damage. The results showed that IC50 of chlorogenic acid, ferulic
acid, orientin and isoorientin was 2 726.11, 1 680.28, 1 889.15 and 2 358.56 μmol/L, respectively. At experimental doses
of IC50, 1/2 IC50, 1/4 IC50, and 1/8 IC50 for these four monomers, the change of mitochondrial membrane potential was in
a dose-dependent manner. Meanwhile, ferulic acid resulted in the largest decease in mitochondrial membrane potential,
followed by chlorogenic acid, orientin and isoorientin. The production of ATP was negatively correlated with the dose of
flavonoids, which was inhibited by ferulic acid, orientin, chlorogenic acid and isoorientin in this decreasing order. In the case
of cytochrome C release, the decreasing order was ferulic acid, orientin, isoorientin and chlorogenic acid. In summary, all
the four flavonoids can result in mitochondrial damage at higher or lower levels. Ferulic acid has the highest mitochondrial
toxicity, followed by orientin, isoorientin and chlorogenic acid. The change of mitochondrial function may be considered as
an index of early cellular damage.

Key words: cytochrome C, mitochondrial membrane potential, adenosine triphosphate, mitochondrial damage, chlorogenic acid, ferulic acid, orientin, isoorientin