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太白楤木皂苷的抗氧化作用研究

 育1,杨 沛2,周雪莹2,海春旭2,秦绪军2,*,李金钢1,*   

  1. 1. 陕西师范大学生命科学学院,陕西 西安 710062;2. 第四军医大学预防医学院毒理学教研室,陕西 西安 710032
  • 收稿日期:2013-04-09 修回日期:2013-06-12 出版日期:2013-07-30 发布日期:2013-07-30
  • 通讯作者: 秦绪军,E-mail:qinxujun@hotmail.com; 李金钢,E-mail:jingang@snnu.edu.cn
  • 作者简介: 郭 育 (1987- ),女,陕西省西安市人,硕士研究生,研究方向:动物行为生理生态。
  • 基金资助:

    国家自然科学基金项目 (81270417);陕西省中医药管理局课题 (zy35)

The antioxidative activity of saponin from Aralia taibaiensis

GUO Yu1YANG Pei2,ZHOU Xue-ying2,HAI Chun-xu2,QIN Xu-jun2,*,LI Jin-gang1,*   

  1. 1. School of Life Science, Shaanxi Normal University, Xi’an 710062; 2. Department of Toxicology, School of Preventive Medicine, the Fourth Military Medical University, Xi’an 710032, Shaanxi, China
  • Received:2013-04-09 Revised:2013-06-12 Online:2013-07-30 Published:2013-07-30
  • Contact: QIN Xu-jun,E-mail:qinxujun@hotmail.com; LI Jin-gang,E-mail:jingang@snnu.edu.cn

摘要:

目的: 探讨太白楤木皂苷的抗氧化作用,为进一步利用其防治疾病提供基础数据。方法:采用体外化学法检测太白楤木皂苷的总抗氧化能力。在细胞实验中,采用氧化剂叔丁基过氧化氢 (tert-butyl hydroperoxide,tBHP)诱导细胞氧化损伤。设立相应对照组和药物保护组,给予太白楤木皂苷 (5 μg/mL)预处理保护,收集样品,流式细胞仪荧光法检测细胞活性氧 (reactive oxygen species,ROS)水平,Western blot方法检测抗氧化转录因子核呼吸因子2 (nuclear respiratory factor 2,Nrf2)及其下游的血红素加氧酶1 (heme oxygenase-1,HO-1)和谷氨酸半胱氨酸连接酶 (γ-glutamate-cysteine ligase,GCL)的表达水平。结果:太白楤木皂苷体外实验检测总抗氧化能力表现出明显的剂量-效应关系。在细胞实验中,tBHP可以显著提高细胞ROS水平,引起细胞的氧化损伤,太白楤木皂苷预处理可以显著降低tBHP诱导的细胞ROS水平。同时发现太白楤木皂苷可以显著拮抗tBHP引起Nrf2及其下游的HO-1、GCL 的催化亚基 (GCLC)和调节亚基 (GCLM)表达水平的升高。结论:太白楤木皂苷在化学反应体系及细胞水平,均显现出了抗氧化作用,具有抗氧化防治疾病的应用前景。

关键词: 太白楤木, 抗氧化, 活性氧, 皂苷, 核呼吸因子2

Abstract:

 OBJECTIVE: To evaluate the antioxidative effect of saponin from Aralia taibaiensis,and provide some basic data for its further application in the prevention and treatment for oxidative stress-related diseases. METHODS: The total antioxidant capacity of saponin was determined by ferric-reducing ability of plasma (FRAP) method. In the cellular study,tert-butyl hydroperoxide (tBHP) was employed as an oxidant to induce oxidative stress,and saponin (5 μg/mL) was pre-incubated with the cells 12 h before challenged by tBHP in the protective group. Then cellular ROS was labeled with DCFH-DA and determined by flow cytometry. The protein levels of nuclear respiratory factor 2 (Nrf2),heme oxygenase-1 (HO-1),γ-glutamate-cysteine ligase (GCL) were evaluated by Western blot. RESULTS:The saponin showed marked antioxidative effect. tBHP induced massive reactive oxygen species (ROS) production,for which saponin pre-treatment showed significant inhibition. Saponin pre-treatment also significantly inhibited the tBHP-stimulated protein expressions of Nrf2,HO-1 and GCL. CONCLUSION:Saponin from Aralia taibaiensis has potent antioxidative activity,indicating its potential clinical application in oxidative stress-related diseases.

Key words:  Aralia taibaiensis, antioxidative, reactive oxygen species, saponin, nuclear respiratory factor 2