3种抑郁症大鼠模型代谢组学的比较  

Comparison of Metabolomics in Three Depression Rat Models

作　　者：刘萌萌 路静 李唯嘉 张文静 陈鸿雁 王雨辰 林贺[1] 林喆[1] 律广富[1] LIU Mengmeng;LU Jing;LI Weijia;ZHANG Wenjing;CHEN Hongyan;WANG Yuchen;LIN He;LIN Zhe;LYU Guangfu(Changchun University of Chinese Medicine,Changchun 130117,China;Affiliated Hospital of Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学,长春130117 [2]长春中医药大学附属医院,长春130117

出　　处：《中国实验方剂学杂志》2024年第16期149-159,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：吉林省科技发展计划项目(20220505038ZP)。

摘　　要：目的:通过比较抑郁状态,以及脑、肝、肾组织中的代谢物差异,评价3种不同方式建立的抑郁症大鼠模型,为抑郁症动物模型的制备和抑郁症标准化诊断、治疗及研究提供参考。方法:40只健康雄性大鼠,随机分为空白组(K组)、慢性不可预知组(GY组)、肾虚型抑郁症组(SM组)和脾虚型抑郁症组(PM组)。K组正常饲养;GY组采用慢性不可预知性温和刺激(CUMS)联合孤养造模;SM组采用CUMS联合皮下注射氢化可的松(25 mg·kg^(-1))孤养造模;PM组采用CUMS联合腹腔注射利血平(0.5 mg·kg^(-1))孤养造模,造模时间均持续4周。采用行为学实验评估动物抑郁程度;酶联免疫吸附测定法(ELISA)检测大鼠血清中5-羟色胺(5-HT)、去甲肾上腺素(NE)的水平;采用超高效液相色谱-四级杆-飞行时间串联质谱法(UPLC-Q-TOFMS)分别对各组大鼠的脑、肝脏、肾脏组织进行代谢组学分析,并进行主成分分析(PCA)和正交偏最小二乘法判别分析(OPLS-DA),通过变量重要性投影(VIP)值>1筛选差异代谢物,并进一步通过京都基因与基因组百科全书(KEGG)数据库对各组大鼠脑、肝、肾组织进行通路富集分析。结果:大鼠行为学结果显示,与K组比较,3种模型组大鼠的糖水消耗率均显著下降(P<0.01);强迫游泳实验中的静止时间显著增加、攀爬次数显著下降(P<0.01);在旷场实验中,运动距离减少,角落活动时间及静止时间增加(P<0.05,P<0.01),GY组、SM组中心区域活动时间显著降低(P<0.01),PM组中心区域活动时间明显增加(P<0.05);在社会交互实验中,与Stranger1接触时间显著减少(P<0.01),与Familiar及Stranger2接触时间明显减少(P<0.05,P<0.01);Morris水迷宫实验中的逃避潜伏期显著增加,逃逸平台滞留时间、平台象限进入次数及平台穿台次数显著下降(P<0.01);血清5-HT、NE水平均明显降低(P<0.05,P<0.01)。代谢组学结果显示,3种模型组大鼠的脑、肝、肾组织代谢物与代谢通路均�Objective:The rat models of depression established in three different ways were evaluated by comparing the depressive state,and the differences of metabolites in brain,liver and kidney tissues,in order to provide a reference for the preparation of animal models of depression and the standardized diagnosis,treatment and research of depression.Method:Forty healthy male rats were randomly divided into blank group(K group),chronic unpredictable group(GY group),kidney deficiency depression group(SM group)and spleen deficiency depression group(PM group).K group was reared normally,GY group was modeled by chronic unpredictable mild stimulation(CUMS)in combination with solitary rearing,and SM group was modeled by CUMS combined with subcutaneous injection of hydrocortisone(25 mg·kg^(-1))and solitary rearing,group PM was modeled by CUMS combined with intraperitoneal injection of rifampicin(0.5 mg·kg^(-1)),and the modeling period lasted for 4 weeks in all groups.Behavioral experiments were used to evaluate the degree of depression,the levels of 5-hydroxytryptamine(5-HT)and norepinephrine(NE)in serum of rats were detected by enzyme-linked immunosorbent assay(ELISA).The metabolomics of brain,liver and kidney tissues of rats in each group were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS),and principal component analysis(PCA)and orthogonal partial least squaresdiscriminant analysis(OPLS-DA)were performed to screen the differential metabolites by variable importance in the projection(VIP)value>1,and pathway enrichment analysis was performed by Kyoto Encyclopedia of Genes and Genomes(KEGG)database.Result:The behavioral results showed that compared with the group K,the sugar water consumption rate of the three model groups was significantly decreased(P<0.01).In the forced swimming test,the rest time increased significantly and the climbing times decreased significantly(P<0.01).In the open field experiment,the movement distance decreased,the corner activity ti

关 键 词：抑郁症 中医证候模型 代谢组学 差异代谢物 模型评价 氢化可的松 利血平 超高效液相色谱-四级杆-飞行时间串联质谱法(UPLC-Q-TOF-MS) 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学] R969.1R749