青藏高原黄刺多糖精细结构解析及其干预胰岛β细胞凋亡作用机制研究

黄刺(Berberis dasystachya M.)作为一种青藏高原特色浆果资源，在民间食用和药用历史悠久。前期研究发现黄刺粗多糖具抗氧化、抗肿瘤及降血糖活性，可显著改善Ⅱ型糖尿病大鼠胰岛β细胞凋亡。本项目拟在前期研究基础上解析黄刺均一多糖的化学结构，揭示其干预胰岛β细胞凋亡的分子机制，并探讨可能存在的降血糖作用机制。首先利用红外光谱、紫外光谱、液相色谱、质谱、气质联用、核磁共振、扫描电镜、原子力显微镜等近现代仪器解析均一多糖的理化性质、一级结构及高级结构；其次利用STZ、高糖及高脂诱导的NIT-1胰岛β细胞凋亡模型，研究其干预胰岛β细胞凋亡的活性；最后构建动物实验模型，从整体水平考察其对胰岛细胞凋亡相关指标的影响，并利用分子生物学技术在细胞和分子水平研究其干预胰岛细胞凋亡的分子机制。项目为寻找多糖核心活性片段及多糖构效关系研究提供科学依据，为青藏高原黄刺多糖的开发利用奠定理论指导基础。

As a featured berry resources in Qinghai-Tibet Plateau, Berberis dasystachya M. has a long history of medicinal and edible utilization in the folk. Based on our previous studies that polysaccharides of Berberis dasystachya M. (BDPs) exhibited antioxidant, antitumor and hyperglycemia activities, it also could significantly interven islet β cell apoptosis in typeⅡ diabetic rats. On the basis of our previous work, the chemical structure of homogeneous polysaccharide from Berberis dasystachya M. (BDPH) will be analyzed, the intervening effect of homogeneous polysaccharide on NIT-1 pancreatic β cells apoptosis and its molecular mechanisms can be revealed, and the protective mechanisms against hyperglycemia will be explored in this study. Firstly, the chemical structure of BDPH will be analyzed by the modern instruments such as IR spectra, UV spectra, liquid chromatography (LC), mass spectrometry (MS), GC-MS combination, nuclear magnetic resonance (NMR), scanning electron microscope (SEM) and atomic force microscope (AFM), etc. Secondly, the intervening effect of BDPH on NIT-1 pancreatic β cells damaged by streptozotocin (STZ), high glucose(HG), high fat(HF) will be interpreted in vitro, respectively. Lastly, The animal experiment models are planed to be constructed to discuss the effects of the BDPH on the pancreatic β cells apoptosis related indexes on global level, and the effects of BDPH on the pancreatic β cells associated factors must be studied through the molecular biology technology on the cellular and molecular level. Furthermore, such findings may supply valuable and fundamental information to further develop on seeking active polysaccharide fragment and clarifying structure- hyperglycemia activity relationship. This research will also provide certain theoretical guidance for the comprehensive utilization on polysaccharides of Berberis dasystachya M. in Qinghai-Tibet Plateau.