指纹图谱和体外细胞活性跟踪相结合考察温度对灵芝中三萜类成分的影响

目的:探讨温度对灵芝中主要抗肿瘤活性三萜类成分含量的影响.方法:采用HPLC灵芝醇提取物中三萜类成分在不同温度处理后的指纹图谱的变化,并结合体外人肺癌SPC-A-1细胞株,通过MTT法评价不同温度影响下的灵芝提取物对肿瘤细胞增殖的抑制作用.结果:灵芝中三萜类成分显著受到温度的影响.在选定的20个特征峰中,第4,14,15,16号峰的峰面积变化经100℃加热处理后最为显著,与未加热处理的灵芝提取物的色谱峰相比,这4个峰的峰面积下降幅度分别高达42.601％,35.909％,49.359％,64.061％.体外细胞活性实验表明经不同温度处理下的灵芝提取物的抗肿瘤活性不同,灵芝提取物的IC50由1.44 g·L-1(醇提取物)升高到2.77 g·L-1(100℃下处理的样品).结论:温度可显著影响灵芝抗肿瘤活性三萜类成分的含量变化,高温易造成其活性成分的损失,进而引起灵芝抗肿瘤活性的降低.

Studies on the Influence of Temperature on the Stability of Triterpenes in Herba Ganoderma lucidum by HPLC and Anti-tumor Activity Assays in vitro

Objective:To study the influence of temperature on the stability of the triterpenes in Ganoderma lucidum.Method: Investigating the differences in the HPLC fingerprints of the ethanol extract of G.lucidum treated at different temperatures,and SPC-A-1 cells were used to evaluate the inhibitory effects in vitro of these different extracts on tumor cell growth by MTT assays.Result: Triterpenes contents of G.lucidum could be significantly affected by temperature.In the 20 selected chromatographic peaks,the area changes of peak 4,14,15,16 were most obvious.Compared with unheated ethanol extract,the descending range of the four peak areas in the ethanol extract treated at 100 ℃ were 42.601%,35.909%,49.359%,64.061%,respectively.Results in vitro showed that the ethanol extract treated at different temperatures had different anti-tumor activities.The IC50of unheated ethanol extract was 1.44 mg crude drug·mL-1,however,after the ethanol extract treated at 100 ℃,the IC50 increased to 2.77 mg crude drug·mL-1.Conclusion: Results indicated that temperature could significantly affect the contents of the triterpenes in G.lucidum.High temperature could easily lead to the loss of its active components,thus decreasing its anti-tumor activity.