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目的优选出破壁灵芝孢子粉中总三萜的最佳提取工艺,并对其体外抗肿瘤活性进行评价。方法正交试验法优化灵芝总三萜的提取工艺;MTT法测定体外抗肿瘤活性;流式细胞仪检测筛选出的细胞株的凋亡率。结果最佳提取工艺条件为95%乙醇,液料比60∶1,在85℃条件下,回流提取2 h,提取2次,总三萜的含量可达6.45%;灵芝总三萜对人结肠癌HCT116细胞、人肺癌A549细胞、人乳腺癌MDA-MB-231细胞的IC50值分别为1.29,1.68,3.81 mg·mL~(-1);不同浓度的灵芝总三萜(0.64,1.6,4.0,10 mg·mL~(-1))作用于结肠癌HCT116 36 h能显著诱导结肠癌HCT116细胞发生凋亡。结论该提取工艺提取效率较高,稳定、可行;灵芝总三萜在体外具有抗人结肠癌、人乳腺癌及人肺癌作用,并可显著诱导HCT116细胞产生凋亡作用。
Objective To optimize the extraction of total triterpenoids from broken spores of Ganoderma lucidum and evaluate the antitumor activity in vitro. Methods The orthogonal experiment was used to optimize the extraction of total triterpenoids from Ganoderma lucidum. The antitumor activity of Ganoderma lucidum total triterpenoids was determined by MTT assay. The apoptosis rate of the selected cell lines was detected by flow cytometry. Results The optimal extraction conditions were 95% ethanol, liquid to liquid ratio 60: 1, reflux extraction for 2 h at 85 ℃, extraction twice, total triterpene content up to 6.45% The IC50 values of HCT116 cells, human lung cancer A549 cells and human breast cancer MDA-MB-231 cells were 1.29,1.68 and 3.81 mg · mL -1, respectively. The total triterpenoids (0.64, 1.6, 4.0) , 10 mg · mL -1) for 36 h in colon cancer HCT116 could induce apoptosis of HCT116 cells. Conclusion The extraction process has the advantages of high extraction efficiency, stability and feasibility. Ganoderma lucidum total triterpenoids have anti-human colon cancer, human breast cancer and human lung cancer effects in vitro and can significantly induce the apoptosis of HCT116 cells.