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目的 探讨脂多糖(lipopolysaccharide)诱导细胞因子紊乱抑郁症模型小鼠海马组织蛋白质组学的变化,研究细胞因子紊乱所致抑郁症的相关发病机制.方法 根据糖水消耗基线值和体重将60只健康雄性Charles River Cancer Research CD-I(R)(ICR)小鼠采用随机数字表法分为实验组(腹腔注射无菌脂多糖0.83 mg/kg)和对照组(腹腔注射无菌生理盐水1 ml/kg),每组30只.提取2组小鼠的海马组织蛋白,采用传统的双向凝胶电泳和基质辅助激光解析电离飞行时间串联质谱进行差异蛋白质组学分析,采用免疫印迹法进行蛋白验证.结果 (1)行为学评价:与对照组比较,实验组小鼠24 h液体消耗量(t=-8.529,P=0.008)和糖水偏好(t=-9.463,P=0.006)均降低,强迫游泳实验不动时间(t=3.105,P=0.003)和悬尾实验不动时间(t=3.228,P=0.002)延长,提示建模成功.(2)双向凝胶电泳和质谱分析:共得到47个在实验组表达上调和59个在实验组表达下调的蛋白质.差异蛋白质主要参与谷氨酸/γ-氨基丁酸代谢、能量代谢、突触传递、细胞骨架构成等生物学过程.(3)能量代谢相关蛋白单磷酸腺苷依赖性蛋白激酶(adenosine 5’-monophosphate-activated protein kinase,AMPK)免疫印迹法验证结果显示,与对照组比较,实验组pAMPK-α(0.84±0.17与1.24±0.33)、pAMPK-β (0.66±±0.38与1.53±0.45)、AMPK-γ2 (0.39±0.10与0.60±0.17)呈下调趋势,差异有统计学意义(t=-2.662,-3.125,-2.491,P=0.024,0.005,0.032);2组AMPK-α和AMPK-β的差异无统计学意义.结论 脂多糖诱导的细胞因子紊乱模型抑郁症小鼠海马组织的蛋白质组学可发生明显改变,AMPK表达水平明显降低,提示发病机制可能与能量代谢障碍有关.“,”Objective To explore the differential proteomics of hippocampal tissue proteins and the pathogenesis in a mice depression model which induced by lipopolysaccharide (LPS).Methods According to the baseline of sucrose consumption and weight,60 male Charles River Cancer Research CD-I(R) (ICR) mice were randomly divided into depression model group and control groups (n=30 per group).After establishment of mice depression model by intraperitoneal injection of LPS,the tissue proteins were extracted from hippocampus of both groups.Subsequently,the differential proteomics analysis wasperformed by conventional two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS),and Western blotting was used to protein verification.Results (1)The experimental results from the 24 h sucrose consumption test showed consistent decreases of liquid intake (t=-8.529,P=0.008) and sucrose preference (t=-9.463,P=0.006) in the model group,forced swimming test immobility (t=3.105,P=0.003) and tail suspension test immobility (t=3.228,P=0.002) results showed obvious depressive behavior in the model group,suggesting that the mice depression model had been successfully established.(2)By 2-DE combined with mass spectrometry analysis (2DE-MS),comparing with the control group mice,47 up-regulated and 59 down-regulated proteins in the experimental group were discovered.These proteins were mainly classified into four functional categoriesincluding glutamate/γ-aminobutyric acid (GABA) metabolism,energy metabolism,synaptic transmission,cytoskeleton formation.(3)The expression level of adenosine monophosphate activated protein kinase (AMPK) which related energy metabolism was significant lower in depression model group,it was consistent with the results of 2-DE verified by Western blotting.The average grey value (-x± s) of AMPK-α 、pAMPK-α、A MPK-γ2,AMPK-β,pAMPK-β in model group relatively were (1.05±0.22),(0.84±0.17),(0.39±0.10),(0.16± 0.05),(0.66±0.38),in control group relatively were (0.98±0.14),(1.24±0.33),(0.60±0.17),(0.24±0.11),(1.53 ±0.45).The expression of pAMPK-α(t=-2.662,P=0.024),AMPK-γ2 (t=-2.491,P=0.032),and pAMPK-3(t=-3.125,P=0.005) were significant lower in model group.Conclusion The proteomics of hippocampus could significantly change and expression level of AMPK may be significant lower in cytokines disorder of depression model mice,which indicates that the pathogenesis of cytokines disorder of depression may be associated with energy metabolism disorder.