沙棘叶多酚的分离纯化及其活性分析  

Purification and Activity Analysis of Polyphenols from Hippophae rhamnoides Leaves

作　　者：武海丽 杜锦娥 路洋洋 李汉卿[1,2] 张晟 李卓玉 WU Haili;DU Jin'e;LU Yangyang;LI Hanqing;ZHANG Sheng;LI Zhuoyu(College of Life Science,Shanxi University,Taiyuan 030006,China;Shanxi Key Laboratory for Research and Development of Regional Plants,Taiyuan 030006,China;Institute of Biotechnology,The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education,Shanxi University,Taiyuan 030006,China)

机构地区：[1]山西大学生命科学学院,山西太原030006 [2]特色植物资源研究与利用山西省重点实验室,山西太原030006 [3]山西大学生物技术研究所,化学生物学与分子工程教育部重点实验室,山西太原030006

出　　处：《山西大学学报（自然科学版）》2023年第1期236-243,共8页Journal of Shanxi University(Natural Science Edition)

基　　金：国家自然科学基金(31800290,82072718,31770382);山西省高等学校科技创新项目(2019L0027)。

摘　　要：近年来,药食同源植物因其绿色环保引起了人们极大的关注。山西省是沙棘资源大省之一,但其开发利用远远不足。沙棘叶中富含黄酮、多酚、萜类等化合物,具备较好的保健与医药价值。因此,本研究优化了沙棘叶中自由酚、结合酚的提取,并对其抑制肿瘤细胞生长活性进行评价。首先比较了80%丙酮、80%乙醇、超纯水、80%甲醇、80%乙酸乙酯5种溶剂提取了沙棘叶自由酚的效率,结果发现沙棘叶甲醇提取物中总酚、黄酮、单宁含量均显著高于其他溶剂,且较其它4种溶剂的提取物,其对结肠癌细胞表现出显著的增殖抑制作用。然后采用不同的水解方法从甲醇提取自由态多酚后的沉淀物中提取沙棘叶结合酚,结果表明碱水解法得到的总酚、黄酮、单宁含量及对肿瘤细胞生长抑制活性显著高于酸水解法,且高于自由酚。为得到较纯的活性多酚,进一步分别选用极性、非极性的大孔树脂进行纯化,筛选出最优树脂为HPD600。然后用30%、60%、95%的乙醇梯度洗脱多酚,发现60%的乙醇洗脱液具较强抑制肿瘤细胞增殖的作用。本研究的实施为我省野生沙棘资源在生物医药领域的开发利用提供了理论依据。In recent years, medicinal and edible homologous plants have attracted great attention. Shanxi Province is one of the provinces with large Hippophae rhamnoides resources, but its development and utilization are far from enough. Hippophae rhamnoides leaves are rich in flavonoids, polyphenols and terpenoids, which have good health care and medical value. In this study, the extraction of free phenol and conjugated phenol from the leaves of Hippophae rhamnoides was optimized, and its anti-tumor cell growth activity was evaluated. Firstly, the free phenols of Hippophae rhamnoides leaves were extracted with 80% acetone, 80% ethanol, ultra-pure water, 80% methanol and 80% ethyl acetate. It was found that the total phenol, flavonoid and tannin contents in the methanol extract of Hippophae rhamnoides leaves were significantly higher than the other solvents, and it showed a significant proliferation inhibitory effect on colon cancer cells compared with the other four solvent extracts. Then different hydrolysis methods were used to extract Hippophae rhamnoides leaf-bound phenols from the precipitate after methanol extraction of free state polyphenols. The results showed that the total phenol, flavonoid, tannin content and tumor cell growth inhibitory activity obtained by alkaline hydrolysis were significantly higher than those extracted by acid hydrolysis and those of free phenols. Furthermore, the bound phenols were purified by different polarity macroporous resins, and HPD600 was selected as the optimal resin. Then the polyphenols were sequentially eluted with 30%, 60% and 95% ethanol, and 60% ethanol eluent was found to have a strong inhibitory effect on the proliferation of tumor cells. In this study, polyphenols with growth inhibitory effect on tumor cells were isolated from the leaves of Hippophae rhamnoides, which provided a theoretical basis for the development and utilization of local wild Hippophae rhamnoides resources in the field of biomedicine.

关 键 词：沙棘叶 自由态多酚 结合态多酚 抑制肿瘤细胞活性 

分 类 号：Q26[生物学—细胞生物学]