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目的:分离蟾皮活性组分并考察大孔树脂不同洗脱部位对肺癌细胞A549生长的抑制作用。方法:采用静态吸附-洗脱试验筛选大孔树脂型号,单因素试验优选蟾皮活性组分的大孔树脂纯化工艺。建立乙醇洗脱液中总生物碱和4种蟾毒二烯内酯类成分的含量测定方法,通过MTT试验测定不同洗脱部位对肺癌细胞A549的抑制率。结果:AB-8型大孔树脂对蟾皮总生物碱和4种二烯内酯类成分均具有较好的吸附分离性能,4个不同乙醇洗脱部位对肺癌细胞A549均表现出抑制作用,30%乙醇和70%乙醇洗脱液的细胞抑制率较高,IC50分别为1.83,2.23 mg·L-1。结论:大孔树脂可用于分离纯化蟾皮中活性组分,不同洗脱部位对肺癌细胞A549的抑制率呈现一定的浓度依赖性,30%乙醇和70%乙醇洗脱部位可作为蟾皮抗肿瘤制剂的重点有效组分。
OBJECTIVE: To separate the active components of toad skin and investigate the inhibitory effect of macroporous resin on the growth of lung cancer A549 cells at different elution sites. Methods: The macroporous resin model was screened by static adsorption-elution test. The macroporous resin purification process of the active component of toad skin was optimized by single factor experiment. To establish a method for the determination of total alkaloids and four kinds of bufalodiprolactones in ethanol elution solution, and determine the inhibition rate of lung cancer cell A549 by different elution sites by MTT assay. RESULTS: AB-8 macroporous resin had good adsorption and separation properties for total alkaloids and four diene lactones. Four different ethanol elution sites showed inhibitory effects on A549 cells, The cell inhibition rates of 30% ethanol and 70% ethanol eluent were higher with IC50 of 1.83 and 2.23 mg · L -1, respectively. CONCLUSION: Macroporous resin can be used to separate and purify the active components of toad skin. The inhibitory rate of A549 cells in different elution fractions is concentration-dependent. The elution fraction of 30% ethanol and 70% ethanol can be used as anti-tumor The key active ingredient of the preparation.