| 摘要: |
| 研究酶法提取海红果中总黄酮的最佳工艺及海红果黄酮粗提物对人宫颈癌HeLa细胞增殖作用。采用单因素试验考察纤维素酶、果胶酶、复合酶(不同质量比的纤维素酶和果胶酶)及酶解时间、酶解温度、酶解液pH对海红果总黄酮提取率的影响,运用二次通用旋转回归组合设计优化酶法提取海红果总黄酮的最佳提取工艺参数;采用MTT法探讨海红果黄酮粗提物对人宫颈癌HeLa细胞增殖的影响。结果表明:4个参试因素对海红果中总黄酮提取率影响的大小顺序为:酶的质量浓度>酶解时间>酶解温度>pH。经试验验证,0.25 mg/mL的复合酶(质量比为2:1的纤维素酶和果胶酶)、酶解时间150 min、酶解温度50℃、初始酶解液pH 6.0总黄酮提取率最高,达(0.356 3±0.000 6)%(n=5),与理论计算值0.360 0%相对误差仅1.028%,两者有很好的一致性。海红果黄酮粗提物对HeLa细胞的增殖有较强的抑制作用,其对HeLa细胞增殖的半数抑制浓度(IC50)为109.89 mg/L。酶法提取海红果中总黄酮的提取率比溶剂浸提法有显著提高;海红果黄酮可以作为一种安全有效的抗肿瘤药物。 |
| 关键词: 海红果 黄酮 酶法提取 二次通用旋转回归组合设计 HeLa细胞 |
| DOI：10.11841/j.issn.1007-4333.2013.01.018 |
| 投稿时间：2012-05-15 |
| 基金项目:国家自然科学基金项目(40471053); 陕西省教育厅项目(12JK0647); 陕西省重点学科自然地理学资助 |
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| Effects of extracting technology of total flavonoids in Malus micromalus Makino by enzymic treatment and its flavonoid crude extract on proliferation of Hela cells in vitro |
|
XU Yu-xia, WANG Hua-bin
|
| (Key Laboratory of Disaster Monitoring and Mechanism Simulating of Shaanxi Province, Baoji University of Arts and Science, Baoji 721013, China) |
| Abstract: |
| The study focuses on the technology that can effectively extract total flavonoids from Malus micromalus Makino through enzymatic treatment,and on the investigation of the effects of flavonoid crude extract from Malus micromalus Makino on proliferation of HeLa cells in vitro.The four parameters,enzymatic concentration,treatment duration time,treatment temperature and pH value of enzyme solution were investigated respectively in four experiments,so as to get their respective impacts on the rate of total flavonoid extraction.The parameters of enzymatic treatment for total flavonoids from Malus micromalus Makino was optimized by the quadratic general rotation combination design.The influence of inhibitory action of flavonoid crude extract from Malus micromalus Makino on HeLa cells growth was also investigated by MTT assay.The results showed that the influencing sequence of the four parameters is enzymatic concentration,duration,and pH value of enzyme solution.And the optimized values of above four parameters for the maximum yield of total flavonoids are:0.25 mg/mL (the ratio of cellulase to pectinase is 2:1(g/g)),150 min,50℃,and pH 6.0,respectively.Under the optimized conditions,the experimental value of yield is (0.356 3±0.000 6)% (n=5),the experimental values agreed well with the estimated value of 0.360 0% with relative error 1.028%.Flavonoid crude extract from Malus micromalus Makino shows a strong inhibitory action on growth of HeLa cells in Vitro,with IC50 value of 109.89 mg/L.Compared with the traditional direct alcohol extract process,the optimized enzyme process can observably increase the extracting amount of total flavonoids. |
| Key words: Malus micromalus Makino flavonoids enzymatic extraction quadratic regression general rotary unitized design Hela cells |
