四角菱茎多酚的超声提取工艺优化及体外抗肿瘤活性研究

摘要

OBJECTIVE:To optimize the ultrasonic extraction technology of polyphenol from the stem of Trapa natans and conduct in vitro antitumor activity test. METHODS:Using the yield of polyphenol as dependent variable,ethanol concentration,ex-traction time and liquid-material ratio as independent variables,the ultrasonic extraction technology of polyphenol from the stem of T. natans was optimized by response surface methodology designed by Box-Behnken design. The effects of different concentrations of polyphenol from the stem of T. natans on the proliferation of HeLa cells,U251 cells and HepG2 cells were detected by MTT as-say,IC50 were calculated,and morphologic changes of them were observed. RESULTS:The optimal condition as follows as the liq-uid-material ratio was 34,45% ethanol,ultrasonic extracting for 24 min at 50 ℃. Under this condition,the yield of polyphenol was 29.36 mg/g(RSD=0.72%,n=3),the relative error of which from predicted value 29.89 mg/g was 1.77%. The proliferation of HeLa cells,U251 cells and HepG2 tumor cells were all significantly inhibited by polyphenol with IC50 of 166.2,141.7,126.6μg/ml,showing good dose-effect relationship. The apoptosis of those cells was presented to certain extent. CONCLUSIONS:Opti-mized ultrasonic extraction technology of polyphenol from the stem of T. natans by response surface methodology is simple and pre-dictable. The polyphenol from the stem of T. natans has significant anti-tumor effect in vitro.%目的：优化四角菱茎中多酚的超声提取工艺并考察其体外抗肿瘤活性。方法：以多酚得率为因变量，以乙醇体积分数、提取时间、液料比为自变量，采用Box-Behnken设计的响应面法优化四角菱茎中多酚的超声提取工艺；采用MTT法检测不同质量浓度的四角菱茎多酚对人宫颈癌HeLa细胞、人胶质瘤U251细胞、人肝癌HepG2细胞体外增殖的影响，计算其半数抑制浓度（IC50），并观察细胞的形态变化。结果：最优超声提取工艺为液料比34，45％乙醇，在50℃下超声提取24 min。在此优化条件下多酚得率为29.36 mg/g（RSD＝0.72％，n＝3），与模型预测值29.89 mg/g的相对误差为1.77％。四角菱茎多酚对HeLa、U251、HepG2细胞均具有明显的增殖抑制作用，IC50分别为166.2、141.7、126.6μg/ml，并呈现良好的量效关系，且各细胞均出现一定程度的凋亡形态。结论：响应面法优化所得四角菱茎多酚超声提取工艺方法简单、预测性良好；四角菱茎多酚具有较好的体外抗肿瘤活性。