目的 整合腦組織代謝組學(xué)和腸道菌群測序分析研究酸棗仁湯（Suanzaoren Decoction，SZRD）抗抑郁的作用機制。方法 30只雄性SD大鼠隨機分為對照組、模型組、文拉法辛（0.035 g/kg）組和SZRD低、高（12、24 g/kg）劑量組。除對照組外，其余各組大鼠均進行慢性不可預(yù)知溫和應(yīng)激（chronic unpredictable mild stress，CUMS）造模，同時ig相應(yīng)藥物，連續(xù)給藥28 d?；诔咝б合嗌V-串聯(lián)質(zhì)譜（ultra performance liquid chromatography-tandem mass spectrometry，UPLC-MS/MS）技術(shù)進行腦組織神經(jīng)遞質(zhì)測定。基于超高效液相色譜-四極桿飛行時間質(zhì)譜（ultra performance liquid chromatography-quadrupole time of flight mass spectrometry，UPLC-Q-TOF-MS）技術(shù)進行腦組織非靶向代謝組學(xué)研究，通過多元統(tǒng)計對代謝輪廓進行分析，以變量重要性投影值（variable importance in projection，VIP）＞1、P＜0.05篩選差異代謝物，并通過MetaboAnalyst進行通路分析。采用腸道菌群測序（16S ribosomal RNA，16S rRNA）技術(shù)對腸道菌群進行結(jié)構(gòu)表征，并用氣相色譜-質(zhì)譜聯(lián)用技術(shù)（gas chromatography-mass spectrometry，GC-MS）對短鏈脂肪酸含量進行測定。將差異菌群的相對豐度，短鏈脂肪酸含量和差異代謝物的相對含量與行為學(xué)結(jié)果進行Spearman分析。結(jié)果 SZRD可顯著升高腦中的5-羥色胺（5-hydroxytryptamine，5-HT）和谷氨酸（glutamic acid，Glu）的含量（P＜0.01）。腦代謝組學(xué)結(jié)果顯示，與對照組比較，模型組中共鑒定到43個差異代謝物，其中8個脂肪酸和3個氨基酸相對含量顯著降低（P＜0.05），4個磷脂酰膽堿，4個磷脂酰乙醇胺和1個鞘脂相對含量顯著升高（P＜0.05）；與模型組比較，SZRD低、高（12、24 g/kg）劑量組分別能顯著回調(diào)15、11個差異代謝物（P＜0.05），共同顯著回調(diào)了10個差異代謝物（P＜0.05），且共同作用于苯丙氨酸、酪氨酸和色氨酸生物合成，丙氨酸、天冬氨酸和谷氨酸代謝和三羧酸循環(huán)（tricarboxylic acid cycle，TCA）3條代謝通路。同時，SZRD可顯著升高乳酸桿菌和糞球菌的豐度（P＜0.05）并顯著增加丙酸，乙酸和丁酸的含量（P＜0.05）。Spearman分析表明，乳酸桿菌等差異菌的豐度及乙酸、丁酸、丙酸的含量與抑郁行為密切相關(guān)，差異代謝物的相對豐度與抑郁行為也顯著相關(guān)。結(jié)論 SZRD發(fā)揮抗抑郁作用機制可能與回調(diào)乳酸桿菌、糞球菌、乙酸、丙酸、丁酸及丙氨酸、天冬氨酸和谷氨酸代謝等通路有關(guān)。

Objective To investigate the anti-depression mechanism of Suanzaoren Decoction (酸棗仁湯, SZRD) by integrated brain metabolomics and gut microbiota sequencing analysis. Methods A total of 30 male SD rats were randomly divided into control group, model group, venlafaxine (0.035 g/kg) group, SZRD low- and high-dose groups (12, 24 g/kg). Except control group, chronic unpredictable mild stress (CUMS) modeling was performed and drugs were ig administered for 28 d. Determination of neurotransmitters in brain tissue based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technique was conducted. Non-targeted metabolomic study of brain tissue was conducted based on ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) technology, metabolic profile was analyzed by multivariate statistics, differential metabolites were screened by variable importance in projection (VIP) > 1 and P < 0.05, and pathway analysis was performed by MetaboAnalyst. The gut microbiota was characterized by 16S ribosomal RNA (16S rRNA) and the content of short-chain fatty acids was determined by gas chromatography-mass spectrometry (GC-MS). The relative abundance, short-chain fatty acid content, and metabolite content of different bacteria were compared with behavioral results by Spearman analysis. Results SZRD could significantly increase the content of 5-hydroxytryptamine (5-HT) and glutamic acid (Glu) in brain (P < 0.01). Brain metabolomics results showed that compared with control group, a total of 43 differential metabolites were identified in model group, specifically, the relative contents of eight fatty acids and three amino acids were significantly reduced (P < 0.05), and the relative contents of four phosphatidylcholine, four phosphatidylethanolamine and one sphinspolipid were significantly increased (P < 0.05). SZRD low- and high-dose groups could significantly reduce 15 and 11 differential metabolites (P < 0.05), respectively. All of them significantly reversed ten different metabolites (P < 0.05), and together acted on three pathways of phenylalanine, tyrosine and tryptophan biosynthesis, alanine, aspartate and glutamate metabolism and tricarboxylic acid cycle (TCA). At the same time, SZRD could significantly increase the abundance of lactobacillus and faecalis (P < 0.05) and significantly increase the contents of propionic acid, acetic acid and butyric acid (P < 0.05). Spearman analysis showed that the abundance of Lactobacillus and other differential bacteria and the contents of acetic acid, butyric acid and propionic acid were closely correlated with depressive behavior, and the relative abundance of differential metabolites was also significantly correlated with depressive behavior. Conclusion SZRD can exert antidepressant effect. The mechanism of the antidepressant effect may be related to the regulation of lactobacillus, fecal bacteria, acetic acid, propionic acid, butyric acid, alanine, aspartate and glutamate metabolism pathway.

R285.5

山西省科學(xué)技術(shù)廳-地產(chǎn)中藥功效物質(zhì)研究與利用山西省重點實驗室項目（201605D111004）;山西省科學(xué)技術(shù)廳山西省科技創(chuàng)新人才團隊項目（202304051001020）;山西省科技成果轉(zhuǎn)化引導(dǎo)專項計劃項目（202204021301063）;山西省科學(xué)技術(shù)廳中央引導(dǎo)地方科技發(fā)展資金項目（YDZJSX2021C025）;山西中醫(yī)藥大學(xué)科技創(chuàng)新團隊項目（2022TD2009）;山西中醫(yī)藥大學(xué)2024年科技創(chuàng)新能力培育計劃項目（2024PY-JL-11-01，2024PY-JL-11-02）