目的 探究益腎養(yǎng)心安神片(YYA)通過對應(yīng)激大鼠下丘腦-垂體-腎上腺(HPA)軸調(diào)控進而改善學(xué)習(xí)記憶能力的作用。方法 SPF級SD大鼠隨機分別為對照組、模型組、艾司唑侖片(ES,陽性對照,0.6 mg·kg^-1)、安神補腦液(AS,陽性對照,2 mL·kg^-1)和YYA高、中、低劑量(7.2、3.6、1.8 g·kg^-1)組,采用咖啡因和環(huán)磷酰胺ip同時聯(lián)合水環(huán)境法制作大鼠應(yīng)激模型,造模成功后ig給藥,每天1次,連續(xù)給藥28 d。實驗結(jié)束后采用Morris水迷宮分析學(xué)習(xí)記憶能力的變化;HE染色觀察海馬區(qū)病理形態(tài)學(xué)改變;實時熒光定量PCR(qRT-PCR)法和免疫組化法檢測海馬區(qū)血管活性腸肽(VIP)和神經(jīng)生長因子(NGF)的表達;ELISA法檢測腦組織超氧化物歧化酶(SOD)活力的變化、血清促腎上腺皮質(zhì)激素(ACTH)、腎上腺皮質(zhì)激素釋放激素(CRH)和腎上腺皮質(zhì)激素(ACH)以及腦組織中5-羥色胺(5-HT)含量。結(jié)果 與模型組比較,各給藥組大鼠穿越平臺和在目標象限探索的時間明顯增加,其中ES組和YYA中、高劑量組尤為顯著(P＜0.05);海馬區(qū)神經(jīng)元病變程度明顯改善;海馬區(qū)神經(jīng)元細胞胞漿內(nèi)NGF和VIP染色明顯增強,NGF和VIP mRNA表達水平顯著升高(P＜0.05、0.01);ES組、YYA中、高劑量組SOD水平顯著升高(P＜0.01);各給藥組血清中ACTH和CRH水平均顯著降低(P＜0.05、0.01),AS組、YYA高劑量組ACH水平顯著降低(P＜0.05);AS組、ES組、YYA高劑量組5-HT水平顯著升高(P＜0.05、0.01)。結(jié)論 YYA可以通過抑制HPA通路,降低腦組織氧化應(yīng)激水平,從而改善海馬組織損傷,營養(yǎng)保護神經(jīng),進而增強應(yīng)激大鼠的學(xué)習(xí)記憶能力。

Objective To explore the pharmacological mechanism of Yishen Yangxin Anshen(YYA) Tablets by axial regulation of the hypothalamic-pituitary-adrenal pathway on the model of stress rats.Methods SPF grade SD rats were randomly divided into control group, model group, Eszolam tablets(ES, positive control, 0.6 mg·kg^-1), Anshen Bu Nao Ye(AS, positive control, 2 mL·kg^-1),and YYA high, medium, and low dose(YYA 7.2, 3.6, 1.8 g·kg^-1) groups. The model of stress rats was made by intraperitoneal injecting cyclophosphamide and caffeine and water environment. After successful modeling, ig administration was administered once a day for 28 consecutive days. Morris Water Maze was used to detect learning memory ability. HE staining was used to observe the pathological changes in the hippocampus. Immunohistochemistry and RT-PCR were applied to detect the expression of nerve growth factor(NGF) and vasoactive intestinal peptide(VIP) in the hippocampus. ELISA was used to detect the level of serum adrenocorticotropic hormone(ACTH), adrenocorticoid-releasing hormone(CRH), adrenal corticoid hormone(ACH), and surperoxide dismutase(SOD), 5-hydroxytryptamine(5-HT) content in the brain tissue.Results Compared with the model group, the time for rats to cross the platform and explore in the target quadrant significantly increased in each treatment group, with the ES group and YYA medium and high dose groups being particularly significant(P＜0.05). The degree of neuronal damage in the hippocampus has significantly improved. The staining of NGF and VIP in the cytoplasm of hippocampal neurons was significantly enhanced, and the expression levels of NGF and VIP mRNA were significantly increased(P＜0.05, 0.01). The SOD levels in the ES group, YYA medium and high-dose groups significantly increased(P＜0.01). The levels of ACTH and CRH in the serum of each treatment group were significantly reduced(P＜0.05, 0.01), while the levels of ACH in the AS group and YYA high-dose group were significantly reduced(P＜0.05). The levels of 5-HT were significantly increased in the AS group, ES group, and YYA highdose group(P＜0.05, 0.01).Conclusion YYA can reduce the oxidative stress level of the brain tissue, reduce the hippocampal tissue damage, protect nerves, and enhance the learning and memory ability of the stressed rats by inhibiting the HPA pathway.

河北省高層次人才資助項目(E2020100001)