首都医科大学学报 ›› 2011, Vol. 32 ›› Issue (3): 371-374.

• 基础研究 • 上一篇    下一篇

褐藻多糖硫酸酯对组织蛋白酶B活性影响的实验研究

张甘霖1,朱晓新2,李萍3*   

  1. 1. 北京中医药大学中药学院,北京 100029; 2. 中国中医科学院中药研究所,北京 100700; 3. 北京市中医研究所,北京 100010
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-06-21 发布日期:2011-06-21
  • 通讯作者: 李萍

Fucoidan alleviates apoptosis by inhibiting cathepsin B and oxidative stress

ZHANG Gan-lin1, ZHU Xiao-xin2, LI Ping3*   

  1. 1. School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100029, China;2. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;3. Institute of Traditional Chinese Medicine, Beijing 100010, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-06-21 Published:2011-06-21
  • Contact: LI Ping

摘要:

目的 研究褐藻多糖硫酸酯(Fucoidan)对氧化应激损伤及溶酶体组织蛋白酶B活性的影响,探讨Fucoidan用于中枢神经系统疾病的作用机制。
方法 50 μg·L-1神经生长因子(nerve growth factor,NGF)分化大鼠肾上腺嗜铬细胞瘤PC12细胞(differentiated PC12,dPC12)7 d,0.5 mmol/L H2O2刺激30 min,给予10 mg·L-1 Fucoidan干预。酶标仪检测细胞内超氧化物歧化酶(superoxide dismutase,SOD)及活性氧(reactive oxygen species,ROS)活性;荧光底物法检测细胞质中溶酶体组织蛋白酶B的活性;酶-底物法检测不同浓度Fucoidan(0.1、1、10、50、100 mg·L-1)对人肝脏溶酶体组织蛋白酶B活性的影响。
结果 氧化损伤后细胞中ROS显著升高达正常水平的185.2%,伴有SOD下降,溶酶体组织蛋白酶B活性上升至正常水平的236.7%;Fucoidan对SOD活性没有明显影响,能降低ROS水平,且对细胞质内和人肝脏溶酶体组织蛋白酶B的活性均具有抑制作用。
结论 Fucoidan降低活性氧含量抗氧化损伤的作用可能与其抑制溶酶体组织蛋白酶B的活性有关。

关键词: 褐藻多糖硫酸酯, 溶酶体组织蛋白酶B, 阿尔茨海默病, 氧化应激

Abstract:

Objective To study mechanism of Fucoidan in neurodegenerative disease by observing its effect on cathepsin B activities and oxidative stress.
Methods After 7 days’ differentiation by 50 μg·L-1 nerve growth factor(NGF), PC12 cells were exposed to 0.5 mmol/L H2O2 for 30 min, and then treated with Fucoidan(10 mg·L-1). The activities of superoxide dismutase(SOD) and reactive oxygen species(ROS) inside cells of each group were assayed. Cathepsin B activities inside cells were determined after being extracted from dPC12. Human liver cathepsin B activities were tested after being exposed to different concentrations of Fucoidan(0.1, 1, 10, 50, 100 mg·L-1).
Results After oxidative stress, the activities of ROS inside cells increased to 185.2% of normal control, accompanied by lower SOD activities and higher activities of cathepsin B(236.7%) in cytoplasm. Fucoidan treatment decreased ROS levels without any effect on SOD; furthermore, Fucoidan could inhibit both dPC12 cytoplasm cathepsin B activities and human liver cathepsin B activities.
Conclusion Fucoidan’s suppression on cathepsin B activities contributes to its protective effect against oxidative stress which leads to cell apoptosis.

Key words: Fucoidan, cathepsin B, Alzheimer&rsquo, s disease, oxidative stress

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