雪松松针总皂苷的超声提取工艺优选及体外抗肿瘤活性研究

摘要

OBJECTIVE:To optimize the ultrasonic extraction technology of total saponins from pine needles of Cedrus deoda-ra(TSPNCD),and to investigate its in vitro antitumor activity. METHODS:The content of TSPNCD was determined by UV spec-trophotometry. Based on single factor test,ethanol volume fraction,solvent-material ratio and extraction time were selected as influ-ential factors during ultrasonic extraction. According to Box-Behnken design,using the content of TSPNCD as response value,re-sponse surface analysis was conducted and used to optimize the ultrasonic extraction technology of TSPNCD. The effects of TSPNCD on the proliferation of human lung cancer A549 cells,human hepatocellular carcinoma HepG2 cells and human gastric cancer MKN45 cells were detected,and IC50 of them were calculated. RESULTS:Optimal technology was that extraction solvent was 73% ethanol;solvent-material ratio was 13∶1;ultrasonic extraction times was twice,each time for 65 min. Under the above conditions,the average content of TSPNCD reached 6.693 mg/g,and relative error between it and predicted value (6.508 mg/g) was 2.84%. TSPNCD inhibited the growth of A549 cells,HepG2 cells and MKN45 cells in a dose-dependent manner,with the IC50 values of (63.98 ± 6.79),(154.91 ± 10.20),(176.32 ± 14.26) µg/ml,respectively. CONCLUSIONS:The ultrasonic extraction technology was verified as reasonable and feasible. TSPNCD has certain inhibitory effect on the proliferation of A549 cells,HepG2 cells and MKN45 cells,especially for A549 cells.%目的：优选雪松松针总皂苷的超声提取工艺，并考察其体外抗肿瘤活性。方法：采用紫外分光光度法测定雪松松针总皂苷的含量。在单因素试验基础上，以乙醇体积分数、液料比、提取时间为因素，根据Box-Behnken设计进行试验，以雪松松针总皂苷含量为响应值进行响应面分析，对雪松松针总皂苷的超声提取工艺进行优化。考察所制雪松松针总皂苷对人肺癌A549细胞、人肝癌HepG2细胞和人胃癌MKN45细胞增殖的影响，并计算半数抑制浓度（IC50）。结果：优选工艺为乙醇体积分数73％，液料比13∶1，提取2次、每次65 min；以此工艺所得雪松松针总皂苷平均含量为6.693 mg/g ，与预测值6.508 mg/g的相对误差为2.84％。雪松松针总皂苷对A549、HepG2、MKN45细胞的增殖均有抑制作用，且呈明显的浓度依赖性，IC50分别为（63.98±6.79）、（154.91±10.20）、（176.32±14.26）µg/ml。结论：优选的雪松松针总皂苷的超声提取工艺合理、可行；其对A549、HepG2、MKN45细胞增殖均有一定的抑制作用，其中对A549细胞作用最为明显。