产前应激致抑郁样子代大鼠海马磷酸化蛋白质和代谢产物变化的初步研究

Phosphor-proteomics and metabolomics in hippocampus of prenatal stress induced depression-like behavior of offspring rats

目的探讨产前应激(PS)诱导抑郁样子代大鼠大脑海马磷酸化蛋白质和代谢产物的基本特征,为儿童抑郁研究提供分子机理。方法本研究将怀孕母鼠随机分为对照组(n=8)和PS组(n=8),PS组在妊娠第14~20 d接受慢性束缚应激,使用糖水偏好实验(SPT)对45只出生后30 d的雄性子代大鼠进行PS易感性(PS-S)筛选;进一步随机选取PS-S组和对照组各8只,采用旷场实验(OFT)、强迫游泳实验(FST)、悬尾实验(TST)评价PS模型的有效性;应用串联质谱标签(TMT)标记的磷酸化蛋白质组学和基于超高效液相色谱-四极杆飞行时间质谱联用技术的非靶向代谢组学,定量分析PS组和对照组子代大鼠之间磷酸化蛋白质和代谢产物的差异。结果与对照组相比,PS-S子代大鼠在OFT的穿越总格数、中心格数、直立次数和在中央格内的停留时间显著减少(t值分别为14.70、6.396、9.058、14.27,P<0.05),在FST和TST中的不动时间显著延长(t值分别为17.70、8.751,P<0.05);本研究结果显示PS-S组和对照组之间存在197个显著差异磷酸化肽段(140个丰度升高,57个丰度降低),80个显著差异代谢产物(55个水平升高,25个水平降低);不同的磷酸化肽段和代谢产物之间存在相关性;联合分析结果表明,差异修饰蛋白和代谢产物共同参与了胆碱能突触、甘油磷脂代谢、长时程增强等通路。结论本研究系统分析了PS-S子代大鼠海马独特的蛋白质磷酸化谱及代谢产物谱,揭示了差异分子在PS诱导子代大鼠抑郁样行为过程中的生物学功能,以及磷酸化蛋白质与代谢产物之间的调控作用。

Objective To explore the characteristics of phosphorylated proteins and metabolites of the hippocampus in prenatal stress(PS)induced depression-like behavior of male offspring rats,and to provide molecular mechanisms for the study of childhood depression.Methods The pregnant female rats were randomly divided into control(n=8)and PS group(n=8).Rats in PS group were subjected to chronic restraint stress on the 14 th-20 th day of pregnancy.The susceptibility to PS(PS-S)of 45 male offspring rats aged 30 days after birth was screened by the sucrose preference test(SPT).Eight each for control and PS-S group were randomly selected for further evaluation of the validity of animal model by open field test(OFT),forced swim test(FST),tail suspension test(TST).Using tandem mass tag based phosphorylated proteomics and untargeted metabolomics based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry,this study quantitative analyzed of the differences in phosphorylated proteins and metabolites between the PS group and the control group offspring rats.Results Compared with the control group,across the total grid number,across the center grid number,the number of vertical erectile and the time spent in the central grid in OFT were all significantly reduced in PS-S offspring rats(t=14.70,6.396,9.058 and 14.27,respectively,P<0.05).The immobility time in FST and TST was all significant prolonger in PS-S group than in control group(t=17.70 and 8.751,respectively,P<0.05).There were 197 significantly differential phosphor-peptides(140 up-regulated and 57 down-regulated),and 80 significantly differential metabolites(55 up-regulated and 25 down-regulated)between the PS-S group and the control group.There were correlations between different phosphorylated peptides and metabolites.The results of joint analysis showed that differentially modified proteins and metabolites jointly participated in cholinergic synapses,glycerophospholipid metabolism,long-term potentiation and other pathways.Conclusion Our study systematically analyzed the unique profiles of phosphor-peptides and metabolites of the hippocampus in PS-S offspring rats,to uncover the biological function of differential molecular in PS-induction process and regulation of phosphorylated protein to metabolite relationships.