目的 研究提取鮑姆纖孔菌中總三萜的最佳工藝條件,比較鮑姆纖孔菌子實體總三萜和菌絲體總三萜的體外抗乳腺癌細胞(MCF-7)活性.方法 在單因素的基礎上運用Box-Behnken中心組合設計,以超聲溫度、超聲時間、乙醇濃度3個因素3水平的實驗模型,建立提取回歸方程.以響應曲面法(RSM)對實驗結果進行分析,對鮑姆纖孔菌總三萜提取工藝進行優(yōu)化;用CASY細胞分析儀檢測細胞活力,TUNEL法檢測細胞凋亡,分析鮑姆纖孔菌子實體總三萜和菌絲體總三萜抑制乳腺癌細胞增殖和促進乳腺癌細胞凋亡活性.結果 修正最優(yōu)條件為超聲時間40 min,乙醇體積分數(shù)為70%,超聲溫度為60 ℃,此條件下總三萜的最高提取率為9.83 mg/g.驗證實驗結果平均提取率為9.81 mg/g,與模型預測值較為接近;鮑姆纖孔菌子實體總三萜在0.2 mg/mL時,菌絲體總三萜在0.4 mg/mL時,對MCF-7細胞的增殖有顯著的抑制,抑制效果呈濃度相關性.鮑姆纖孔菌子實體總三萜在0.2 mg/mL時,菌絲體總三萜在0.4 mg/mL時,能顯著增強MCF-7細胞的凋亡,增強效果呈濃度相關性.結論 曲面響應法可以對鮑姆纖孔菌總三萜的提取工藝進行優(yōu)化;鮑姆纖孔菌三萜對乳腺癌細胞(MCF-7)有明顯的抑增殖活性和凋亡誘導作用,呈濃度相關性,且子實體總三萜比菌絲體總三萜體外抗乳腺癌細胞(MCF-7)活性更強.

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.