基于生物信息学技术探索丙戊酸盐导致大鼠孤独症样行为的机制  

Mechanism of valproate-induced autism-like behavior in mice based on bioinformatics technique

作　　者：金柳荫 夏玉洁 徐文倩 舒畅[1] 王高华[1] Jin Liumeng;Xia Yujie;Xu Wenqian;Shu Chang;Wang Gaohua(Mental Health Center of Renmin Hospital,Wuhan University,Wuhan 430000,China)

机构地区：[1]武汉大学人民医院精神卫生中心,430000

出　　处：《神经疾病与精神卫生》2023年第2期125-132,共8页Journal of Neuroscience and Mental Health

摘　　要：目的 利用生物信息学技术探索丙戊酸盐导致Wistar大鼠出现孤独症样行为的具体机制。方法 选取SPF级Wistar雄性大鼠4只(300~350 g),雌性大鼠8只(200~250 g),于8~10周将雌、雄大鼠以2︰1的比例合笼过夜。在雌鼠怀孕的第12.5天随机选取4只腹腔注射600 mg/kg的丙戊酸盐,产下的幼鼠纳入丙戊酸盐组,并于幼鼠出生后35 d通过旷场实验、高架十字迷宫实验、新物体识别实验和埋珠实验评价造模是否成功。另随机选取4只孕鼠注射等量生理盐水一次,产下的幼鼠纳入健康对照组。使用GSE42904数据集寻找服用丙戊酸盐和服用甲基纤维素的小鼠脑组织中表达有差异的基因,采用基因本体论(GO)、京都基因与基因组百科全书(KEGG)、GO/KEGG(联合LogFC)以及基因探针富集分析(GSEA)对差异基因进行富集分析。利用实时定量聚合酶链式反应(q-PCR)验证差异基因在丙戊酸盐组幼鼠脑组织中的表达情况。最终纳入丙戊酸盐组幼鼠14只,健康对照组幼鼠7只。结果 在旷场实验中,丙戊酸盐组的中心区累积时间、身体伸直次数少于健康对照组,在新物体识别实验中的识别指数低于健康对照组,埋珠实验中的埋珠数量多于健康对照组,差异有统计学意义(P<0.05)。在高架十字迷宫实验中,丙戊酸盐组与健康对照组进入开放臂次数、运动总距离比较,差异无统计学意义(P>0.05)。提示造模成功。对GSE42904数据集进行分析,符合|Log2(FC)|>1且P<0.05的基因有267个,这些差异基因主要富集于代谢通路。对谷胱甘肽代谢通路中的差异基因谷胱甘肽S-转移酶α 3(Gsta 3)、γ-谷氨酰转移酶7(Ggt 7)和葡萄糖-6-磷酸脱氢酶(G6pd)进行验证,结果显示,两组Gsta3、Ggt7基因的mRNA表达水平比较,差异无统计学意义(P>0.05);丙戊酸盐组G6pd基因的mRNA表达水平低于健康对照组,差异有统计学意义(P<0.05)。结论 在丙戊酸盐大鼠模型中的G6pd基因表达存在差异,Objective To explore the mechanism of valproate-induced autism-like behavior in Wistar mice based on bioinformatics technique.Methods A total of 4 SPF grade Wistar male mice(300-350 g)and 8 female mice(200-250 g)were selected.The female and male mice were caged overnight at a ratio of 2︰1 in 8-10 weeks.The female mice were intraperitoneally injected with 600 mg/kg valproate on the 12.5 days of pregnancy.The newborn mice were included in the valproate group.Open field test,elevated cross maze test,new object recognition test and bead embedding experiment were used to evaluate the success of the model at the 35th day after birth.Another 4 pregnant mice were randomly selected to inject the same amount of normal saline once,and the newborn mice were included in the healthy control group.GSE42904 data set was used to search for genes with different expressions in the brain tissues of mice receiving valproate and methylcellulose.Genetic ontology(GO),Kyoto Encyclopedia of Genes and Genome(KEGG),GO/KEGG(combined with LogFC)and Gene Set Enrichment Analysis(GSEA)were used for enrichment analysis of differential genes.Quantitative real-time polymerase chain lock reaction(Q-PCR)was used to verify the expression of differential genes in the brain tissue of model mice in valproate group.Finally,14 young mice in the valproate group and 7 young mice in the healthy control group were included.Results Compared with the control group,the time of entering the central area and the number of body extensions decreased in the open field experiment,the recognition index decreased in the new object recognition experiment,and the number of buried beads increased in the bead embedding experiment,and all the differences were statistically significant(P<0.05).In the elevated cross maze test,there was no statistically significant difference between the valproate group and the healthy control group in the number of times of entering the open arm and the total distance of movement(P>0.05),indicating successful modeling.According to the analysi

关 键 词：丙戊酸盐 孤独症谱系障碍 葡萄糖-6-磷酸脱氢酶 谷胱甘肽代谢 大鼠 

分 类 号：R749.94[医药卫生—神经病学与精神病学]