目的 研究提取鲍姆纤孔菌中总三萜的最佳工艺条件,比较鲍姆纤孔菌子实体总三萜和菌丝体总三萜的体外抗乳腺癌细胞(MCF-7)活性.方法 在单因素的基础上运用Box-Behnken中心组合设计,以超声温度、超声时间、乙醇浓度3个因素3水平的实验模型,建立提取回归方程.以响应曲面法(RSM)对实验结果进行分析,对鲍姆纤孔菌总三萜提取工艺进行优化;用CASY细胞分析仪检测细胞活力,TUNEL法检测细胞凋亡,分析鲍姆纤孔菌子实体总三萜和菌丝体总三萜抑制乳腺癌细胞增殖和促进乳腺癌细胞凋亡活性.结果 修正最优条件为超声时间40 min,乙醇体积分数为70%,超声温度为60 ℃,此条件下总三萜的最高提取率为9.83 mg/g.验证实验结果平均提取率为9.81 mg/g,与模型预测值较为接近;鲍姆纤孔菌子实体总三萜在0.2 mg/mL时,菌丝体总三萜在0.4 mg/mL时,对MCF-7细胞的增殖有显著的抑制,抑制效果呈浓度相关性.鲍姆纤孔菌子实体总三萜在0.2 mg/mL时,菌丝体总三萜在0.4 mg/mL时,能显著增强MCF-7细胞的凋亡,增强效果呈浓度相关性.结论 曲面响应法可以对鲍姆纤孔菌总三萜的提取工艺进行优化;鲍姆纤孔菌三萜对乳腺癌细胞(MCF-7)有明显的抑增殖活性和凋亡诱导作用,呈浓度相关性,且子实体总三萜比菌丝体总三萜体外抗乳腺癌细胞(MCF-7)活性更强.

Objective To optimize the ultrasonic-assisted extraction technology of total triterpenoids from Inonotus baumii, and compare the inhibitory activities of total triterpenoids from fruiting body and from mycelium on breast cancer cells (MCF-7) in vitro. Methods A three-factor and three-level Box-Behnken experimental design was employed using ultrasonic temperature, ultrasonic time and ethanol concentration as a model. Response surface method (RSM) was used to analyze the result and optimize the extracting process. The anticancer activities of total triterpenoids from fruiting body and from mycelium against MCF-7 were completed by CASY cell analyzer and TUNEL method, respectively. Results Optimum extraction conditions were determined as follows: extraction time of 40 min, concentration of ethanol 70%, and extraction temperature of 60 ℃. Under these conditions, the yield of total triterpenoids reached 9.83 mg/g. The experimental results were verified with an average yield of 9.81 mg/g, which is close to the model predicted value. The concentration of total triterpenoids from fruiting body of I. baumii reached 0.2 mg/mL, and that from mycelium reached 0.4 mg/mL, which could inhibit hyperplasia of MCF-7 in dose-dependent manner. The concentration of total triterpenoids from fruiting body of I. baumii reached 0.2 mg/mL, and that from mycelium reached 0.4 mg/mL, which could significantly enhanced MCF-7 apoptosis in dose-dependent manner. Conclusion The RSM technology optimization for extracting total triterpenoids from I. baumii is suitable. Total triterpenoids from fruiting body and mycelium of I. baumii could inhibit proliferation of MCF-7, and induced apoptosis on tumor cells in dose-dependent manner. Total triterpenoids from fruiting body are more active than that from mycelium against MCF-7.