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

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

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·m L-1、600μg·m L-1、1 200μg·m L-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·m L-1)对肝细胞增殖几乎无抑制作用;中浓度(600μg·m L-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·m L-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·m L-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p H 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。

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, 600μg·mL-1, 1 200μg·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 （300μg·mL-1）. It showed certain inhibition rate in middle concentration group （600μg·mL-1）, but disappearedwhile glutathione added. The inhibition rate decreased significantly with the increment of the amount of GSH added in high concentration group （1 200μg·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.