蜂花粉对酒精性肝细胞氧化应激损伤的保护作用与机制研究

Bee pollen is one of the important products of bee collection. In recent years, the hepatoprotective effect of bee pollen has attracted considerable attention, and the obvious protective effect of bee pollen against liver injury has been shown by animal experiment. However, functional components and molecular mechanism of bee pollen protecting against alcoholic liver injury is not clear. In our preliminary study, the results showed that the phenolic compounds of bee pollen could increase the viability of hepatic cells under alcohol-induced injury. In order to further clarify the effect and mechanism of different phenolic components of bee pollen protecting against alcohol-induced liver injury, on the basis of preliminary study, in this project, the chemical structures of phenolic compounds of bee pollen are identified, and using alcohol-injured hepatic cells as a model, the protective effects of different fractions of phenolic compounds against alcohol-induced oxidative stress injury in hepatic cells are compared and the phenolic fraction possessing the strongest protective effect is obtained. The molecular mechanism of bee pollen protecting against alcohol-induced oxidative stress injury in hepatic cells is investigated from a new perspective that the activation of Nrf2-ARE pathway and promotion of intracellular redox homeostasis recovery. The aim of this project is to provide theoretical basis for the elucidation of substance elements and mechanism of bee pollen protecting against alcoholic liver injury. The implementation of this research will not only provide a new way to broaden the application of bee pollen in food, health products and other fields, but also have a positive practical significance to promote the beekeepers income and the sustainable development of China's apiculture.

蜂花粉是蜜蜂采集行为的重要产品之一，其保肝功效近年来受到广泛关注，动物实验证明蜂花粉对肝损伤具有显著的保护作用，但其对酒精性肝损伤保护的功能因子和分子机制并不清楚。申请人前期研究发现蜂花粉酚类物质能提高酒精损伤下的肝细胞活力。为了进一步明确蜂花粉不同酚类组分对酒精性肝损伤的保护效果和作用机制，本项目拟在前期研究的基础上，对蜂花粉中酚类物质进行鉴定，采用酒精损伤肝细胞模型，比较蜂花粉不同酚类组分对酒精性肝细胞氧化应激损伤的保护作用，筛选得到保护作用最强的组分；进而从活化Nrf2-ARE信号通路，促进细胞内氧化还原稳态恢复的新角度探讨蜂花粉保护酒精性肝细胞氧化应激损伤的分子机制，为阐明蜂花粉保护酒精性肝损伤的物质基础和作用机制提供理论依据。本项目的实施不仅为拓宽蜂花粉在食品、保健品等领域的应用提供新思路，而且对促进蜂农增收，推动我国养蜂业的可持续发展，具有积极的现实意义。

我国的蜂花粉资源丰富，但蜂花粉精深加工产品很少。蜂花粉的保肝活性是其重要活性之一，阐明其保肝活性因子和保肝机制有利于蜂花粉保肝产品的开发。针对上述问题，本项目主要完成了以下研究工作：（1）对五味子蜂花粉、油菜蜂花粉和茶花蜂花粉中的酚类物质进行了分离和鉴定，明确了五味子蜂花粉中的酚类物质主要为酚胺类化合物，油菜蜂花粉和茶花蜂花粉中的酚类物质主要为黄酮类化合物和酚胺类化合物。（2）研究了油菜蜂花粉酚类物质对肝细胞氧化应激损伤的保护作用，证明了油菜蜂花粉中的酚胺类化合物的保肝活性优于黄酮类化合物。（3）研究了五味子蜂花粉酚类物质对酒精性肝细胞氧化应激损伤的保护作用和作用机制，通过酒精损伤HepG2细胞模型，证明了五味子蜂花粉酚类物质对酒精性肝细胞氧化应激损伤具有明显的保护效果，酚胺化合物是蜂花粉保肝活性的重要活性因子；采用qRT-PCR和Western blot技术分析Nrf2-ARE信号通路中的细胞保护蛋白和重要因子，明确了五味子蜂花粉酚类物质通过调控Nrf2-ARE信号通路发挥保肝功效。本项目的研究结果为开发蜂花粉保肝产品提供理论依据，对提高蜂花粉附加值和推动蜂花粉产业发展具有重要意义。