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目的:研究瑞香狼毒总生物碱的提取分离、纯化和含量测定,确定总生物碱成分的体外抗肿瘤作用,通过对细胞周期和凋亡的检测研究抗肿瘤机理。方法:采用酸提碱沉淀法提取总生物碱,用阳离子交换树脂进行纯化,剩余滴定法测定总生物碱含量。体外抗肿瘤活性研究采用MTT比色法和集落形成法测定,靶细胞采用人胃癌细胞SGC-7901、人肝癌细胞BEL-7402和人白血病细胞HL-60,流式细胞仪检测药物作用后肿瘤细胞周期和细胞凋亡情况。结果:提取纯化后瑞香狼毒总生物碱含量为68.24%,总生物碱对体外培养的三株肿瘤细胞有较强的抑制作用,主要机理为导致细胞G0/G1期阻滞,诱导肿瘤细胞凋亡。结论:瑞香狼毒总生物碱提取物具有较强的体外抗肿瘤作用。
OBJECTIVE: To study the extraction, isolation, purification and determination of total alkaloids from Stellera chamaejasmelon, and to determine the antitumor effect of total alkaloids in vitro. To study the antitumor mechanism by detecting cell cycle and apoptosis. Methods: Total alkaloids were extracted by acid-base precipitation and purified by cation exchange resin. The total alkaloids content was determined by titration. The antitumor activity in vitro was determined by MTT colorimetric assay and colony formation assay. The target cells were treated with human gastric cancer cell line SGC-7901, human hepatocellular carcinoma cell line BEL-7402 and human leukemia cell line HL-60. Flow cytometry Cycle and Apoptosis. Results: The total alkaloid content of Stellera chamaejasme was 68.24% after extraction and purification. The total alkaloids had a strong inhibitory effect on the three tumor cells cultured in vitro. The main mechanism was the block of G0 / G1 phase and the induction of tumor cell apoptosis Death. Conclusion: Stellera chamaejas total alkaloids extract has a strong anti-tumor effect in vitro.