蓝莓花青素酰化产物的稳定性及诱导肝癌细胞凋亡的机制研究

项目名称： 蓝莓花青素酰化产物的稳定性及诱导肝癌细胞凋亡的机制研究

项目编号： No.31271981

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 孙爱东

作者单位： 北京林业大学

项目金额： 80万元

中文摘要： 花青素性质不稳定, 易受食品体系中pH、温度和介质的影响。目前常用的花青素稳定化技术包括添加抗氧化剂、微胶囊化 、糊精包合等，但效果并不显著。本项目拟通过在分离、纯化的蓝莓花青素的分子结构上引入没食子酰基, 获得改性产物-花青素酰化物。在探讨光照、pH、温度等因素对酰化物颜色影响的基础上，利用激光光散射法测定花青素酰化物的分子量, 质谱、红外、核磁检测其结构变化。通过建立亚急性大鼠肝癌模型，探索花青素酰化物对肝损伤大鼠的保护和氧化应激作用。并进一步采用改良MTT 法( WST-8法)观察酰化产物对肝癌细胞的生长抑制，碘化丙锭单染色研究细胞周期改变，Annexin V- FITC/PI双染流式细胞术检测细胞凋亡水平，聚焦显微镜观察和Western blotting分析细胞自噬发生。结合花青素酰化物体外清除自由基抗氧化活性，从抗氧化角度探讨其对肝癌细胞的抑制作用及其可能作用机制。

中文关键词： 蓝莓；花青素；酰化；抗氧化；抗癌

英文摘要： The property of anthocyanins were unstable and influenced by the pH, temperature and media.The common technology for stabilization anthocyanin include adding antioxidants,microcapsule,dextrin inclusion,but the effect was not significant.This program was by drawing galloyl acyl group into molecular structure of anthocyanin that seperated and purified from blueberry,to obtain modified products-acylated anthocyanin. On the basis of the effect of light, pH,temperature and other factors on the color of acylated products, using laser light scattering method to determinate the molecular weight of acylated anthocyanin,mass spectrometry, infrared, nuclear magnetic resonance to detect the structural changes. By the establishment of sub-acute rat liver cancer model, to explore the protection and oxidative stress effect of the acylated anthocyanins on liver injury in rats.Using improved MTT assay ( WST-8 method) to further observe the inhibitory action of acylated products to the growth of cell of hepatoma,by propidium iodide staining to detect changes in cell cycle, Annexin V- FITC/PI flow cytometry to detect the level of apoptotic cells, focusing microscope and Western blotting to observe and detect cell autophagy. With antioxidant activities in scavenging free radicals of acylated anthocyanins, discussing the inhibio

英文关键词： blueberry；anthocyanins；acylate；antioxidant；anticancer