目的 探究肉苁蓉苯乙醇苷类成分抗脑缺血再灌注损伤作用的靶点蛋白群及相关作用机制。方法 利用线栓法制备大鼠大脑中动脉阻断模型，考察肉苁蓉苯乙醇苷的脑保护作用。通过构建一种表面键合有肉苁蓉苯乙醇苷的琼脂糖固相微球，利用键合在固相微球表面的苯乙醇苷类分子从脑组织裂解液中特异性捕获相应的脑保护作用相关靶点蛋白，并进行高分辨质谱鉴定。结果 肉苁蓉苯乙醇苷能够显著改善大鼠脑缺血再灌注损伤，包括降低脑缺血面积、降低缺血相关行为学评分及提高脑组织中超氧化物歧化酶（SOD）水平。利用靶点"钩钓"策略共获得HSP90等18个靶点蛋白。针对这些靶点蛋白的生物学功能，将其分为9类，包括抗氧化、离子通道、免疫调节、细胞存活、细胞骨架等。结论 从靶点源头上解释了肉苁蓉苯乙醇苷发挥抗痴呆、缓解疲劳、抗肿瘤、免疫调节等诸多作用的潜在药理机制，进而也展示了一种解析中药复杂成分体系分子药理机制的新方法。

Objective Cistanches Herba is a kind of tonic traditional Chinese medicine with several therapy effects including tonifying kidney-yin, anti-dementia, anti-aging and relaxing bowel. Phenylethanoid glycosides (PhGs) are the major effective components in C. tubulosa. However, there were no further studies on molecular pharmacologic mechanisms due to its complex components and mechanism diversity of action in PhGs till to now. The aim of this study was to investigate the target protein groups and related mechanisms associated with PhGs in anticerebral ischemia-reperfusion injury. Methods The middle cerebral artery occlusion (MCAO) model was established in rats, and the protective effects of PhGs on cerebral ischemia-induced injuries were determined. A kind of solid bead whose surface was cross-linked with PhGs was prepared to capture the target proteins from brain tissue lysates. The target proteins were further identified with LC-MS/MS. Results PhGs significantly inhibited cerebral ischemia-induced injuries by reducing ischemia size and rat behavioral scores and elevated the SOD levels in rat brain tissues. Eighteen target proteins were identified based on "target fishing" strategy and divided into 9 kinds according to their biological functions, including anti-oxidation, ion channel, immunoregulation, cell survival and cytoskeleton, etc. Conclusion These findings reveal the potential pharmacological mechanisms of PhGs in anti-dementia, fatigability alleviating, anti-tumor, immunoloregulation, etc, and also present a promising technology for investigating the complicated pharmacological mechanisms of traditional Chinese medicine.

国家自然科学基金资助项目（81303253，51273012）