目的 基于“腎腦相關(guān)”理論，探究龜齡集對氫化可的松與β淀粉樣蛋白（Aβ）誘導(dǎo)的阿爾茨海默病模型大鼠的作用，并利用代謝組學(xué)的方法探討其相關(guān)作用機(jī)制。方法 采用ip氫化可的松與icv Aβ誘導(dǎo)的阿爾茨海默病模型，選用SD雄性大鼠60只，分為對照組、模型組、假手術(shù)組和龜齡集低、中、高劑量組。行為學(xué)實(shí)驗(yàn)結(jié)束后，麻醉取血、腎上腺、脾、腎、胸腺、睪丸、腦和海馬，計(jì)算腎上腺指數(shù)及腎臟指數(shù)。采用新物體識別實(shí)驗(yàn)、Morris水迷宮實(shí)驗(yàn)檢測藥物干預(yù)對大鼠認(rèn)知功能的影響。酶聯(lián)免疫試劑盒（ELISA）法檢測各組組織上清液中睪酮和皮質(zhì)酮（CORT）含量。蛋白質(zhì)免疫印跡法（Western blotting）法測定大鼠海馬組織中蛋白激酶B（Akt）、磷酸化蛋白激酶B（p-Akt）的表達(dá)水平。利用非靶向代謝組學(xué)方法，篩選出用藥前后潛在代謝差異物并進(jìn)行通路分析，找到與龜齡集治療阿爾茨海默病模型大鼠所涉及的相關(guān)代謝通路。結(jié)果 新物體識別結(jié)果顯示，與對照組相比，模型組新物體辨別指數(shù)（RI）顯著降低（P＜0.05）；與模型組相比，龜齡集高、中劑量組新物體RI顯著升高（P＜0.05）。Morris水迷宮實(shí)驗(yàn)顯示，與模型組相比，龜齡集高、中、低劑量組大鼠在Ⅲ象限路程、Ⅲ象限時(shí)間顯著延長（P＜0.05）。與模型組相比，龜齡集低、中、高劑量組腎臟指數(shù)和龜齡集高劑量腎上腺指數(shù)均顯著升高（P＜0.01）。血清激素水平結(jié)果顯示：與模型組相比，龜齡集高、中、低劑量組CORT顯著降低（P＜0.05）。Western blotting結(jié)果顯示，與模型組相比，p-Akt/Akt龜齡集高、中、低劑量組表達(dá)水平均顯著升高（P＜0.05、0.01）。用藥前后共得到23種血清差異代謝物水平發(fā)生不同程度的回調(diào)，主要涉及代謝途徑為甜菜堿代謝、初級膽汁酸的生物合成、磷脂生物合成等代謝途徑。結(jié)論 龜齡集對氫化可的松與Aβ誘導(dǎo)的阿爾茨海默病模型大鼠可明顯改善認(rèn)知功能障礙，保護(hù)腦組織海馬結(jié)構(gòu)，改善模型大鼠血清中CORT和睪酮水平。還可以通過調(diào)節(jié)阿爾茨海默病模型大鼠的整體代謝水平。

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

山西中醫(yī)藥大學(xué)科技創(chuàng)新能力培育計(jì)劃（2019PY-126）；山西省中醫(yī)藥大學(xué)中藥炮制學(xué)科建設(shè)項(xiàng)目（2018-1）