摘要
目的观察白杨素(ChR)衍生物8-溴-7-甲氧基白杨素(Br MC)对体外培养肝癌HepG2细胞生长的抑制作用;用人结肠腺癌(Caco-2)细胞模型比较白杨素及其衍生物Br MC的首过糖苷化速率,从药动学角度阐明Br MC比ChR抗肿瘤活性强的可能原因,为寻找既具有较强抗肿瘤活性又具有良好的药动学特性的活性新化学实体提供实验依据。方法①MTT比色法测定Br MC对体外培养肝癌Hep-G2细胞和人胚肝L-02细胞增殖的影响,评价其对肝癌细胞作用选择性;②体外培养Caco-2细胞,用Transwell建立单层细胞模型;③评价时间对Br MC和ChR葡萄糖醛酸化产物形成的影响,用β-葡萄糖醛酸苷酶水解葡萄糖醛酸结合物,高效液相色谱(HPLC)检测,计算葡萄糖苷化速率。结果①MTT比色法结果显示,Br MC(3.0、10.0、30.0μmol.L-1)及其先导化合物ChR(30.0μmol.L-1)与阳性对照药物5-氟尿嘧啶(5-FU30.0μmol.L-1)分别处理肝癌HepG2细胞48 h,其增殖相对抑制率分别为31.5%、52.3%、61.2%、51.1%和60.8%;Br MC的效价强度高于其先导化合物ChR,与5-FU相当。Br MC对正常人胚肝L-02细胞的毒性作用(IC50为446.5μmol.L-1)远小于对肝癌HepG2细胞的毒性(IC50为10.3μmol.L-1),对人肝癌HepG2细胞的药物选择指数达43.3;②Caco-2细胞培养21 d后,成功建立细胞单层模型;③Br MC和ChR的葡萄糖苷化产物均随时间延长而增加,但Br MC比ChR的糖苷化速率显著降低。结论以ChR为先导物,选择性引进溴和甲氧基的衍生物Br MC对肝癌的生长抑制作用较ChR明显增强,可能与其葡萄糖醛酸化程度显著降低,药物在肝癌细胞中的蓄积增加有关。
Objective To observe the effect of the derivatives of chrysin（ChR）,8-bromo-7-methoxychrysin（BrMC）,on the cell growth of human hepatoma cell line（HepG2） in vitro,and pharmacokinetically illuminate why BrMC has stronger biological effect than ChR,through comparing the glucuronidation of BrMC with ChR in Caco-2 monolayer model,and find a new chemical entity with good pharmacodynamics as well as good pharmacokinetics.Methods ①MTT assay was used to determine the effect of BrMC on the proliferation of HepG2 cells and L-02 cells and to evaluate the selectivity of action against tumor cells.②Caco-2 cells were cultured in vitro and Caco-2 monolayer model established by Transwell.③We evaluated the effect of time on glucuronic conjunction formation of BrMC and ChR,HPLC examination.Results ①The MTT assay showed that the inhibitory rate in HepG2 cells was 31.5%,52.3%,61.2%,51.1% and 60.8% treated with BrMC（3.0,10.0,and 30.0 μmol·L-1）,ChR（30.0 μmol·L-1） and 5-FU（30.0 μmol·L-1）,respectively for 48 h.The efficacy of BrMC was stronger than that of ChR,and close to 5-FU.The toxic activity of the BrMC in L-02 cells（IC50,446.5 μmol·L-1） was significantly lower than that of HepG2 cells（IC50,10.3 μmol·L-1）.The selective index was 43.3.②The Caco-2 cells were cultured in Transwell for 21 d,and we established the monolayer cell model successfully.③The glucuronic conjunction of BrMC and ChR increased with time,while the glucuronidation formation rate of BrMC descended markedly.Conclusion The toxic activity of BrMC is highly selective to tumor cells in vitro.After modification by introducing bromine and methoxyl in chrysin,the glucuronidation rate of its derivatives（BrMC） decreases significantly.
出处
《中南药学》
CAS
2011年第4期241-245,共5页
Central South Pharmacy
基金
湖南省自然科学基金项目(No.03JJY5009)
