青稞β-(1,3),(1,4)-葡聚糖包裹弹性蛋白酶壳核结构形成机制及其对降血脂作用的影响

Whether the polysaccharide-protein nanoparticles with the spherical core-shell structure, based on their self-assembly function and the same kind of pharmacological activity, could play their active roles at the same time, and furthermore improve their efficacies because of the protection of the active structure of core protein by the polysaccharide shell? The answer of this question decides the research direction of active amphipathic bio-macromolecules in the research field of nano medicine. The present study chooses Hordeum vulgare var. nudum β-glucan and elastase, both of which have self-assembly function and hypolipidemic activity, to be the research materials, and then explores the probability of using the core-shell nanoparticles for novel drugs and drug delivery system, which are made of polysaccharide and protein with the same kind of pharmacological activity. During the research, we will answer the question “how the H. vulgare β-(1→3),(1→4)-D-glucan and elastase form the spherical core-shell structural polysaccharide-protein nanoparticles through the nanogelated glycosylation reaction? ” from two aspects of key reaction process of covalent binding and nano gelation, and furthermore, answer the question “what effects of the nanogelated glycosylation reaction on the hypolipidemic activity of H. vulgare β-–glucan and elastase? ” from the aspect of the effects of the structure characteristics and molecular morphologies on the hypolipidemic activity, such as molecular weight of β-glucan, grafting degree of nanogels, diameter of nanoparticles. The present research will provide the supplemental basic data of β-(1→3),(1→4)-D-glucan for the nano medicine research, and a novel research approach for the research and development of novel drugs and drug delivery system using cereal β-(1→3),(1→4)-D-glucans.

糖蛋白纳米粒子能否以多糖外壳有效保护蛋白质核心的活性结构，协同发挥多糖和蛋白质的药理作用？这个问题的答案决定今后纳米医药领域活性两亲性生物大分子的研究方向。本项目选择具有自组装功能和降血脂活性的青稞β-葡聚糖和弹性蛋白酶为研究材料，探索将同类药理活性的多糖和蛋白质制备成壳核结构纳米粒子用于新型药物或药物传递体系的可行性，从共价结合和纳米凝胶化两个关键反应进程方面研究“β-(1,3),(1,4)-葡聚糖与弹性蛋白酶如何形成球形壳核结构纳米粒子？”；从反应前后组成变化、β–葡聚糖分子量、纳米粒子枝接度、粒径等结构和分子形貌因素对产物降血脂能力影响方面研究“纳米粒子形成过程对二者的降血脂作用产生怎样的影响？”，补充该领域降血脂糖蛋白纳米粒子和β-(1→3),(1→4)结构葡聚糖基础数据，为同类型谷物β-葡聚糖类新型药物的设计研发提供新思路。

糖蛋白纳米粒子能否以多糖外壳有效保护蛋白质核心的活性结构，协同发挥多糖和蛋白质的药理作用？这个问题的答案决定今后纳米医药领域活性两亲性生物大分子的研究方向。本项目选择具有自组装功能和降血脂活性的青稞β-葡聚糖和弹性蛋白酶为研究材料，探索将同类药理活性的多糖和蛋白质制备成壳核结构纳米粒子用于新型药物或药物传递体系的可行性，从共价结合和纳米凝胶化两个关键反应进程方面研究“β-(1,3),(1,4)-葡聚糖与弹性蛋白酶如何形成球形壳核结构纳米粒子？”；从反应前后组成变化、β–葡聚糖分子量、纳米粒子枝接度、粒径等结构和分子形貌因素对产物降血脂能力影响方面研究“纳米粒子形成过程对二者的降血脂作用产生怎样的影响？”，补充该领域降血脂糖蛋白纳米粒子和β-(1→3),(1→4)结构葡聚糖基础数据，为同类型谷物β-葡聚糖类新型药物的设计研发提供新思路。. 研究结果表明，青稞β-葡聚糖包裹弹性蛋白酶的壳核结构纳米粒子形成机制为青稞β-葡聚糖还原端的活泼羟基和弹性蛋白酶的自由氨基形成希夫碱，再经环化形成，N-取代醛糖基胺，经重排形成糖蛋白共价结合物，随后利用二者的自组装功能和共价结合物上β-葡聚糖链的空间位阻，通过热变性凝胶化反应，控制反应条件，形成多糖外壳、蛋白质内核的糖蛋白纳米粒子。此外，青稞β-葡聚糖的分子量对于形成弹性蛋白酶-β-葡聚糖凝胶纳米粒子的粒径、均匀程度及其降血脂活性有着显著影响：随β-葡聚糖分子量增大，凝胶粒子粒径逐渐增大，但其PDI逐渐减小均一化程度增高。3种糖蛋白纳米凝胶粒子中，分子量居中的nanogel Elastase-HBP-D21的降血脂作用最强，可能是其纳米粒径较nanogel Elastase-HBP-D74小，且高级结构稳定性较nanogel Elastase-HBP-D4强等原因导致的。. 综上所述，糖蛋白纳米粒子能够以多糖外壳有效保护蛋白质核心的活性结构，协同发挥多糖和蛋白质的药理作用。因此，接下来本课题组将继续深入探索将同类药理活性的多糖和蛋白质制备成壳核结构纳米粒子用于新型药物或药物传递体系的可行性。