蟾皮活性组分的分离与体外抗肿瘤活性考察

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。

Separation and in vitro Antitumor Activity Investigation of Active Constituents from Toad Skin

Objective: To isolate active constituents from toad skin and investigate inhibition effect of different macroporous resin elution parts on growth of lung cancer cell line A549. Method: Type of macroporous resin was screened by static adsorption-elution test,single factor tests were adopted to optimize separation and purification technology of active constituents from toad skin.Assay methods of total alkaloids and four bufadienolides were established,inhibition rate of different elution parts on lung cancer cell line A549 was determined by MTT assay. Result: AB-8 macroporous absorption resin was selected with a better adsorption separation property for total alkaloids and four bufadienolides from toad skin,four different elution parts exhibited certain inhibition effect on lung cancer cell line A549,30% and 70% ethanol eluent showed stronger inhibition effect with IC₅₀ of 1.83 and 2.23 mg·L^-1,respectively. Conclusion: Macroporous absorption resin could be used for separation and purification of active constituents from toad skin,inhibition effect of different elution parts on lung cancer cell line A549 showed a certain concentration-dependent,30% and 70% ethanol elution parts could be used as key active ingredients in antitumor preparations of toad skin.