中国药物警戒 ›› 2015, Vol. 12 ›› Issue (7): 385-388.

• 基础及临床研究 •    下一篇

二苯乙烯苷对正常人肝细胞增殖的抑制率及影响因素研究

李娜1,2, 马致洁3, 孟雅坤2, 王亚2, 李晓菲2, 涂灿2, 张雅铭2, 牛明2, 赵艳玲4, 肖小河4, 康廷国1, 游云5, 王伽伯2   

  1. 1. 辽宁中医药大学药学院,辽宁 大连 116600;
    2. 解放军302医院全军中医药研究所,北京 100039;
    3. 首都医科大学附属北京友谊医院,北京 100050;
    4. 解放军302医院中西医结合肝病诊疗与研究中心,北京 100039;
    5. 中国中医科学院中药研究所,北京 100700
  • 收稿日期:2015-09-01 修回日期:2015-09-01 出版日期:2015-07-08 发布日期:2015-09-02
  • 通讯作者: 王伽伯,男,博士,助理研究员,中药肝毒性与合理用药研究。E-mail：wjb0128@126.com。
  • 作者简介:李娜,女,在读博士,实验师,中药学。
  • 基金资助:
    国家自然科学基金项目（81373984）; 国家“重大新药创制”科技重大专项课题（2015ZX09501-004-001-008）; 国家公益性行业科研专项课题（201507004-04）; 中国中医科学院客座研究员联合创新研究项目（ZZ070827）; 北京市自然科学基金项目（7152142）

Research of Inhibition Rate and Influential Factors on Normal Human Liver Cell Line Caused by 2,3,5,4-Tetrahydroxyl Diphenylethylene-2-o-glucoside

LI Na1,2, MA Zhi-jie3, MENG Ya-kun2, WANG Ya2, LI Xiao-fei2, TU Can2, ZHANG Ya-ming2, NIU Ming2, ZHAO Yan-ling4, XIAO Xiao-he4, KANG Ting-guo1, YOU Yun5, WANG Jia-bo2   

  1. 1. College of Pharmacy, Liaoning University of Traditional Chinese Medicine, Liaoning Dalian 116600, China;
    2. China Military Institute of Chinese Medicine, 302 Military Hospital, Beijing 100039, China;
    3. Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China;
    4. Integrative Medicine Center, 302 Military Hospital, Beijing 100039, China;
    5. Institute of Chinese Medicine, Beijing 100700, China
  • Received:2015-09-01 Revised:2015-09-01 Online:2015-07-08 Published:2015-09-02

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摘要： 目的 以L-02细胞为载体,考察不同因素 （谷胱甘肽、过氧化氢、温度等）对不同质量浓度的二苯乙烯苷 （300μg·mL-1、600μg·mL-1、1 200μg·mL-1,分别相当于740μmol·L-1、1 480μmol·L-1、2 960μmol·L-1）肝细胞增殖抑制率的影响。方法 培养的人肝L-02细胞分别给以经不同组合方式（与谷胱甘肽或H2O2组合给药）或不同处理方式（与H2O2组合于不同温度蒸干重溶）处理的二苯乙烯苷,采用MTS法检测二苯乙烯苷对人肝L-02细胞的吸光度,并通过公式计算抑制率,比较各组抑制率差异值。结果 低浓度二苯乙烯苷 （300μg·mL-1）对肝细胞增殖几乎无抑制作用;中浓度 （600μg·mL-1）呈现一定抑制作用,加入谷胱甘肽（GSH）后抑制作用消失;高浓度二苯乙烯苷 （1 200μg·mL-1）随着GSH加入量的增加,抑制率显著下降,与阳性对照药野百合碱 （MCT）（1600μg·mL-1,相当于4.92×103μmol·L-1）表现一致。二苯乙烯苷3个浓度对肝细胞增殖的抑制率（6%、25%和82%）有显著性差异,而加入不同体积分数的H2O2（0.5%和1%）对抑制率的影响变化不大。二苯乙烯苷经水溶解并于不同温度（常温、50℃和90℃）及pH 3条件下蒸干,所得样品对L-02细胞增殖的抑制率与未经处理的二苯乙烯苷组比较,没有明显变化。结论 二苯乙烯苷在一定浓度范围内对肝细胞增殖有不同程度的抑制作用,且该作用与氧化或水解关系不大,具体机制有待进一步研究。

关键词: 二苯乙烯苷, 何首乌, 谷胱甘肽, 肝损伤, L-02细胞

Abstract: Objective To investigate the influence of different factors (glutathione, H2O2, temperature, etc.) on inhibition rate of 2,3,5,4-tetrahydroxyl diphenylethylene-2-o-glucoside (TSG) of various concentrations (300 μg·mL-1, 600 μg·mL-1, 1 200 μg·mL-1) on L-02 cell line. Methods Cultured human liver L-02 cell was given styrene glycosides treated in different ways and the absorbance determination was performed with MTS method. The inhibition rate was calculated by the inhibition rate formula. Results There was almost no inhibition rate in low concentration group (300 μg·mL-1). It showed certain inhibition rate in middle concentration group (600 μg·mL-1), but disappeared while glutathione added. The inhibition rate decreased significantly with the increment of the amount of GSH added in high concentration group (1 200 μg·mL-1), which was consistent with that of positive control monocrotaline (MCT). There existed obvious difference in inhibition rate (6%, 25%, 82%) among the three different concentrations of TSG. And there was little influence on inhibition rate of TSG while adding H2O2 of different concentrations (0.5%, 1%). The inhibition rate of TSG dried at different temperature (room temperature, 50℃, 90℃) and pH 3 was almost the same as that of untreated TSG. Conclusion There exists inhibition function for TSG to some extent within certain concentration range to liver cells, which has nothing to do with oxidation or hydrolyzation.

Key words: 2,3,5,4-tetrahydroxyl diphenylethylene-2-o-glucoside, polygonum multiflorum, glutathione, liver injury, L-02 cell line

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