目的 基于“肾脑相关”理论，探究龟龄集对氢化可的松与β淀粉样蛋白（Aβ）诱导的阿尔茨海默病模型大鼠的作用，并利用代谢组学的方法探讨其相关作用机制。方法 采用ip氢化可的松与icv Aβ诱导的阿尔茨海默病模型，选用SD雄性大鼠60只，分为对照组、模型组、假手术组和龟龄集低、中、高剂量组。行为学实验结束后，麻醉取血、肾上腺、脾、肾、胸腺、睾丸、脑和海马，计算肾上腺指数及肾脏指数。采用新物体识别实验、Morris水迷宫实验检测药物干预对大鼠认知功能的影响。酶联免疫试剂盒（ELISA）法检测各组组织上清液中睾酮和皮质酮（CORT）含量。蛋白质免疫印迹法（Western blotting）法测定大鼠海马组织中蛋白激酶B（Akt）、磷酸化蛋白激酶B（p-Akt）的表达水平。利用非靶向代谢组学方法，筛选出用药前后潜在代谢差异物并进行通路分析，找到与龟龄集治疗阿尔茨海默病模型大鼠所涉及的相关代谢通路。结果 新物体识别结果显示，与对照组相比，模型组新物体辨别指数（RI）显著降低（P＜0.05）；与模型组相比，龟龄集高、中剂量组新物体RI显著升高（P＜0.05）。Morris水迷宫实验显示，与模型组相比，龟龄集高、中、低剂量组大鼠在Ⅲ象限路程、Ⅲ象限时间显著延长（P＜0.05）。与模型组相比，龟龄集低、中、高剂量组肾脏指数和龟龄集高剂量肾上腺指数均显著升高（P＜0.01）。血清激素水平结果显示：与模型组相比，龟龄集高、中、低剂量组CORT显著降低（P＜0.05）。Western blotting结果显示，与模型组相比，p-Akt/Akt龟龄集高、中、低剂量组表达水平均显著升高（P＜0.05、0.01）。用药前后共得到23种血清差异代谢物水平发生不同程度的回调，主要涉及代谢途径为甜菜碱代谢、初级胆汁酸的生物合成、磷脂生物合成等代谢途径。结论 龟龄集对氢化可的松与Aβ诱导的阿尔茨海默病模型大鼠可明显改善认知功能障碍，保护脑组织海马结构，改善模型大鼠血清中CORT和睾酮水平。还可以通过调节阿尔茨海默病模型大鼠的整体代谢水平。

Objective To study the effects of Guilingji on hydrocortisone and Aβ induced Alzheimer's Disease model rats, and the mechanism of action explore by metabomics based on the theory of "kidney - brain related".Methods Model of Alzheimer's disease induced by ip hydrocortisone and intraventricular injection of Aβ was used. Sixty male SD rats were selected and divided into control, model, sham operation, and Guilingji low-dose, medium-dose, and high-dose groups. After the behavioral experiment, the blood, adrenal gland, spleen, kidney, thymus, testis, brain, and hippocampus were collected under anesthesia, and the indexes of adrenal gland and kidney were calculated. New object recognition test and Morris water maze test were used to detect the effects of drug intervention on the cognitive function of rats. The levels of testosterone and CORT in supernatant were determined by ELISA. The expression levels of protein kinase B (Akt) and phosphorylated protein kinase B (P-Akt) in hippocampal tissues of rats were determined by Western blotting. Non-targeted metabolomics was used to screen out potential metabolic differences before and after treatment, and pathway analysis was conducted to find the related metabolic pathways involved in Guilingji's treatment of Alzheimer's disease model rats.Results The new object recognition results showed that the new object recognition index (RI) of model group was significantly lower than that of control group (P < 0.05). Compared with model group, RI of new objects in Guiling high and medium dose groups was significantly increased (P < 0.05). Morris water maze test showed that, compared with model group, the distance and time in quadrant Ⅲ of Gulingji high-dose, medium-dose, and low-dose groups were significantly longer (P < 0.05). Compared with model group, kidney index in Guilingji low, medium, and high dose groups were significantly increased (P < 0.01), and adrenal index in Guilingji high dose group were significantly increased (P < 0.01). The results of serum hormone level showed that, compared with model group, CORT of guilingji high-dose, medium-dose, and low-dose groups was significantly decreased (P < 0.05). Western blotting results showed that compared with the model group, the expression levels of p-Akt/AKT in high, medium, and low dose groups were significantly increased (P < 0.05, 0.01). A total of 23 serum metabolites were found to have different levels of callback before and after treatment, mainly involving the metabolic pathways of betaine metabolism, primary bile acid biosynthesis, phospholipid biosynthesis and so on.Conclusion Guilingji can significantly improve the cognitive dysfunction, protect the hippocampal structure of brain tissue, and improve the levels of CORT and T in serum of hydrocortisone and Aβ-induced Alzheimer's disease model rats. It can also regulate the overall metabolic level of Alzheimer's disease model rats.

R285.5

山西中医药大学科技创新能力培育计划（2019PY-126）；山西省中医药大学中药炮制学科建设项目（2018-1）