目的 采用高分辨核磁共振¹H譜（¹H-NMR）代謝組學技術研究慢性溫和不可預知應激（CUMS）抑郁大鼠腦組織中代謝物及代謝通路的變化，探討抑郁癥的發(fā)病機制。方法 12只雄性SD大鼠隨機分為模型組和對照組，采用CUMS對模型組大鼠進行為期4周的造模，進行稱體質量、曠場實驗和糖水偏愛實驗驗證模型是否成功，造模結束后收集大鼠腦組織。采用兩相提取法（甲醇/氯仿/水）對腦組織進行提取，得到水溶性和脂溶性代謝物。應用¹H-NMR技術結合多元統(tǒng)計和代謝通路分析篩選出與抑郁相關的腦內差異代謝物，并構建其代謝通路。結果 行為學數(shù)據顯示，與對照組比較，模型組大鼠的體質量、糖水偏愛率、曠場的穿格數(shù)和直立次數(shù)均顯著降低（P<0.05），顯示了抑郁狀態(tài)。在大鼠腦組織的¹H-NMR圖譜中共指認出35種內源性代謝產物。腦組織中水溶性和脂溶性代謝物主成分分析（PCA）均顯示模型組與對照組分開，與行為學結果一致，表明造模成功；OPLS-DA分析找到9個水溶性差異代謝物和6個脂溶性差異代謝物。就水溶性代謝物而言，與對照組比較，模型組中肌酐、谷氨酰胺、?；撬岷挺?氨基丁酸含量顯著增加（P<0.05、0.01、0.001），丙二醇、谷氨酸、亮氨酸、纈氨酸和賴氨酸含量顯著降低（P<0.05、0.01、0.001）。就脂溶性代謝物而言，與對照組比較，模型組-（CH₂） n和-N （Me₃）₃含量顯著升高（P<0.05、0.001），膽固醇的C18/19甲基、R-CH₃、CH₂OPO₂-、-CH=CH-含量顯著降低（P<0.05、0.01、0.001）。與對照組比較，CUMS造模后5條代謝通路發(fā)生顯著變化：纈氨酸、亮氨酸和異亮氨酸生物合成，?；撬岷蛠喠蚧撬岽x，丙氨酸、天冬氨酸和谷氨酸代謝，糖降解和糖異生通路和丙酮酸代謝。結論 運用¹H-NMR代謝組學技術結合多元統(tǒng)計分析和代謝通路分析，闡明抑郁癥的發(fā)病機制與能量代謝、氨基酸代謝和神經遞質合成等相關。

Objective To study the metabolic changes of brain tissues of chronic unpredictable mild stress (CUMS) rats by 1H-NMR and obtain the potential biomarkers of CUMS rats to investigate the pathogenesis of depression.Methods A total of 12 male Sprague-Dawley (SD) rats were randomly divided into two groups,namely the control and model groups.The CUMS procedure was conducted for four weeks.The weight test,open field experiment and sugar preference experiment were conducted to verify the success of the model.The brain tissues of rats were collected at the end of the procedure.The endogenous metabolites in brain tissues were extracted by MeOH/CHCl₃/H₂O.And the hydrosoluble and liposoluble metabolites were analyzed by ¹H-NMR with different sequences.The differences of metabolites between control group and CUMS group were analyzed by multivariate statistical analysis.The metabolic pathways were constructed.Results Body weight,sucrose preference,ambulation number and rearing number in open-field test of the CUMS rats decreased significantly after 28d procedure as compared with those of the control groups (P<0.05).Also,in the scores plot of principle component analysis (PCA) on brain metabolites,the CUMS-depressed rats were significantly separated with the control rats,suggesting the CUMS model has been built successfully.Thirty-five metabolites were identified in the ¹H-NMR spectra of brain tissues.Nine hydrosoluble differential metabolites and six fat-soluble differential metabolites were found by OPLS-DA analysis.Taking hydrosoluble metabolites into account,the content of creatinine,glutamine,taurine and gamma-aminobutyric acid in the CUMS rats significantly increased (P<0.05),while propanediol,glutamic acid,leucine,valine and lysine were significantly decreased in CUMS model group as compared with the control group (P<0.05,0.01,and 0.001).As far as fat-soluble metabolites are concerned,compared with the control group,the content of-(CH₂) n and-N (Me₃)₃ in the model group increased significantly (P<0.05,0.001),while the content of C18/19 methyl,R-CH₃,CH₂OPO₂-,and-CH=CH-in cholesterol decreased significantly (P<0.05,0.01,0.001).Compared with the control group,there were significant changes in five metabolic pathways:valine,leucine and isoleucine biosynthesis,taurine and sulfite metabolism,alanine,aspartic acid and glutamic acid metabolism,sugar degradation and gluconeogenesis pathway and pyruvate metabolism.Conclusion Using ¹H-NMR metabonomics combined with multivariate statistical analysis and metabolic pathway analysis,the pathogenesis of depression is related to energy metabolism,amino acid metabolism and neurotransmitter synthesis.

國家自然科學基金資助項目（81803962）；國家"重大新藥創(chuàng)制"科技重大專項課題（2017ZX09301047）