目的 探討滋水清肝飲在應(yīng)用脂多糖(LPS)制備的小鼠抑郁模型中的抗抑郁作用和潛在機制。方法 選取60只小鼠按隨機數(shù)字表隨機分為6組:對照組、模型組、鹽酸氟西汀組(陽性藥,10 mg·kg^-1)和滋水清肝飲低、中、高劑量(8.835、17.670、35.340 g·kg^-1)組,給藥14 d后,除對照組ip等量0.9 %的氯化鈉溶液外,其余各組均ipLPS (0.83 mg·kg^-1),LPS注射結(jié)束24 h后,稱小鼠質(zhì)量,并進行懸尾實驗和強迫游泳實驗;通過蘇木素-伊紅(HE)染色觀察小鼠海馬形態(tài)學(xué)改變;免疫熒光染色檢測海馬小膠質(zhì)細胞標志物離子化鈣結(jié)合適配分子1(Iba-1)熒光強度;試劑盒法檢測小鼠血清中超氧化物歧化酶(SOD)活力、丙二醛(MDA)、腫瘤壞死因子-α(TNF-α)、白細胞介素-1β(IL-1β)、白細胞介素-6(IL-6)、5-羥色胺(5-HT)水平及小鼠海馬吲哚胺2,3-雙加氧酶1(IDO1)、犬尿氨酸(KYN)、5-HT水平;Western blotting法檢測小鼠海馬組織中p-核因子κB (NF-κB)、p-細胞外調(diào)節(jié)蛋白激酶(ERK)蛋白表達。結(jié)果 與模型組相比,滋水清肝飲組小鼠體質(zhì)量均有所回升(P<0.05);各給藥組小鼠的懸尾實驗和強迫游泳實驗不動時間顯著縮短(P<0.05、0.01);各給藥組海馬形態(tài)學(xué)轉(zhuǎn)好,海馬小膠質(zhì)細胞標志物Iba-1表達顯著降低(P<0.05);滋水清肝飲中、高劑量組和鹽酸氟西汀組小鼠血清中的5-HT水平顯著增加(P<0.05),各給藥組血清SOD活力顯著增強(P<0.05),血清TNF-α、IL-6、IL-1β、MDA水平顯著降低(P<0.05、0.01);各給藥組小鼠海馬中IDO1、KYN水平顯著下降(P<0.05、0.01),滋水清肝飲中、高劑量組和鹽酸氟西汀組海馬中5-HT水平顯著上升(P<0.05、0.01);各給藥組海馬組織中p-ERK、p-NF-κB蛋白表達顯著降低(P<0.05、0.01)。結(jié)論 滋水清肝飲顯著改善急性LPS小鼠的抑郁樣行為,其機制可能與抑制ERK/NF-κB通路、降低IDO1水平、改善神經(jīng)炎癥有關(guān)。

Objective The aim of this study was to investigate the antidepressant effect and potential mechanism of Zishui Qinggan Yin (ZSQGY) in the LPS-induced depression mice model. Methods A mouse depression model was established by LPS. A total of 60 mice were randomly selected and randomly divided into six groups according to the random number table, namely control group, model group, Flu group (10 mg·kg^-1), ZSQGY low, medium and high dose (8.835, 17.670, 35.340 g·kg^-1) groups. After 14 days of administration, the control group was injected with the same amount of normal saline. The other groups were ip injected with LPS (0.83 mg·kg^-1). After 24 h LPS injection, mice are weighed and the tail suspension test and forced swimming test were performed. The morphological changes of mice hippocampus were observed by HE staining and Nishi staining, and immunofluorescence staining was used to detect the fluorescence intensity of hippocampal microglia marker Iba-1. The activity of superoxide dismutase (SOD), malondialdehyde (MDA), tumor necrosis factor- α (TNF- α), interleukin-1β (IL-1β), interleukin-6 (IL-6) and 5- hydroxytryptamine (5-HT) in serum of mice and the levels of IDO1, KYN, and 5-HT in the hippocampus of mice were detected by kits. The expression of p-NF-κB, p-ERK, and ERK protein level in mouse hippocampus was detected by Western blotting. Results Compared with model group, the body weight of mice in the ZSQGY group increased (P < 0.05). The immobility time in the tail suspension experiment and forced swimming experiment of mice in each treatment group was significantly shortened (P < 0.05, 0.01). The morphology of the hippocampus improved in each treatment group, and the expression of the hippocampal microglial cell marker Iba-1 was significantly reduced (P < 0.05). The levels of 5-HT in the serum of mice in the middle and high-dose groups of ZSQGY and the Flu group were significantly increased (P < 0.05), the activity of SOD in serum was significantly enhanced in each treatment group (P < 0.05), and the levels of TNF-α, IL-6, IL-1β, and MDA in serum were significantly decreased (P < 0.05, 0.01). The levels of IDO1 and KYN in hippocampus were significantly decreased in each treatment group (P < 0.05, 0.01), the level of 5- HT in hippocampus was significantly increased in medium-dose and high-dose ZSQGY group and fluoxetine hydrochloride group (P < 0.05, 0.01). The protein expressions of p-ERK and p-NF-κB in hippocampus were significantly decreased in each treatment group (P < 0.05, 0.01). Conclusion ZSQGY can significantly improve the depression-like behavior in LPS-induced mice and the mechanism may be related to down-regulating ERK/NF-κB signaling pathway and IDO1 level.

R965

陜西省陜西中醫(yī)藥大學(xué)校級科研課題項目(2021GP25);陜西省自然科學(xué)基礎(chǔ)研究計劃(2022JQ-918);國家自然科學(xué)基金項目(82304504)