基于神经血管单元的冰片促进黄芪甲苷和三七总皂苷配伍抗脑缺血作用的研究

Our previous study of the ingredients (components) combination of Chinese materia medica based on the formula combination theory and pharmacological mechanism find that the main active components combination between Astragalus and Panax notoginseng has the potentiation on anti-cerebral ischemia. However, the effective components of Astragalus and Panax notoginseng are difficult to penetrate the blood brain barrier (BBB) because of their large molecular weights and low fat-solubilities. According to the theory of channel tropism of Chinese material medica, we take borneol that “guide the medicine uplink” to prompt the medicine into BBB to combine with Astragaloside IV and Panax notoginseng saponins (PNS), studying that the combination of borneol and Astragaloside IV, PNS promotes the pharmaceutical components into BBB to enhance the anti-cerebral ischemia effects, investigating the mechanisms that borneol motivates the pharmaceutical components to penetrate BBB through drug transport; Furthermore, we research the multi-links and multi-target effects and mechanisms of borneol combined with Astragaloside IV and PNS against cerebral ischemia on account of the protection of the neurovascular unit (NVU). This study is expected to provide a new research idea integrating modern science, technology and traditional Chinese medicine theory, to provide a exemplary reference model for the studies on the ingredients (components) combination of Chinese materia medica and utilizing formula combination theory to study the synergistic effect; In addition, that elucidating the synergistic principle and mechanism of borneol combined with the main effective components of Astragalus and Panax notoginseng will have important scientific significances for further guiding and developing a new Chinese materia medica of the effective ingredients (components).

我们以往基于方剂配伍理论和药理机制的中药有效组（成）分配伍研究模式研究发现，黄芪和三七的有效成分配伍具有增强抗脑缺血的作用。但黄芪和三七的有效成分相对分子量大、脂溶性低，难以透过血脑屏障（BBB）发挥作用。本项目根据中药归经理论，将具有“引药上行”促进药物透过BBB的冰片与黄芪甲苷和三七总皂苷配伍，研究冰片配伍黄芪甲苷和三七总皂苷促药物成分透过BBB增强其抗脑缺血的作用，从BBB药物转运研究冰片促药物成分透过BBB的机制；并从神经血管单元保护研究冰片配伍黄芪甲苷和三七总皂苷抗脑缺血的多环节、多靶点机理。预期本项目的研究，将为中药复方提供一个整合现代科学技术和传统中医理论的全新研究思路，为中药有效组（成）分配伍和利用方剂配伍理论进行协同增效作用研究提供一个示范性研究模式；对冰片配伍黄芪和三七主要有效成分协同增效原理及机制的阐明，为进一步指导研制新型的中药有效组（成）分药物具有重要的科学意义。

本项目根据中医药归经理论，将具有“引药上行”促进药物透过血脑屏障（BBB）的冰片与黄芪甲苷（AST Ⅳ）和三七总皂苷（PNS）配伍，研究冰片配伍AST Ⅳ和PNS的最佳配伍形式、促进药物透过BBB吸收入脑及其机理，并从神经血管单元保护研究冰片配伍AST Ⅳ和PNS抗缺血性脑损伤的整合调节作用。结果发现：冰片、AST Ⅳ、PNS配伍可增强抗脑缺血再灌注损伤的效应，3种药物的有效配伍剂量为冰片7.5 mg/kg+AST Ⅳ 10 mg/kg +PNS 25 mg/kg。冰片与AST IV、PNS合用后，能促进AST IV及人参皂苷Rb1、Rg1 及三七皂苷R1向大脑皮层富集，尤其是向缺血再灌注侧大脑皮层富集与吸收，增强抗缺血性脑损伤的作用，其作用可能与下调BBB中外排蛋白P-gp、MRP-2、MRP-4、MRP-5和相应基因表达，同时上调摄取蛋白OATP-2表达有关；冰片、AST Ⅳ和 PNS 能不同程度降低脑缺血/再灌注后BBB通透性的增加，减轻脑水肿，对抗脑组织损伤，三药合用有增强效应，机制可能与协同抑制缺血后ZO-1、ZO-2、Occludin 表达下调，保护BBB有关；冰片、AST Ⅳ、PNS配伍能够通过促进脑内神经细胞的增殖，修复受损的神经细胞，增强抗脑缺血再灌注损伤的作用，其作用与调控Wnt/β-catenin信号通路有关；冰片配伍AST Ⅳ和PNS对大鼠脑缺血再灌注后神经元损伤具有保护作用，可减轻神经元和微血管损伤，促进微血管新生，其机制可能与激活Notch信号通路有关；冰片配伍AST Ⅳ和PNS对缺血再灌注脑组织具有保护作用，其机制可能是减轻了神经血管单元的主要成分神经元和微血管基底膜的损伤、抑制星形胶质细胞过度活化、抑制水通道开放和保护血脑屏障而减轻脑水肿，从而对神经血管单元发挥保护作用。