基于转录组测序分析砷暴露影响小鼠认知功能的机制  

Analysis of Mechanism of Arsenic Exposure-induced Cognitive Changes in Mice Based on RNA Sequencing

作　　者：原晓红 胡鑫 杨鸣宇 韩明盛 马艳琴[1] Yuan Xiaohong;Hu Xin;Yang Mingyu;Han Mingsheng;Ma Yanqin(College of Life Sciences,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学生命科学学院,太谷030801

出　　处：《生态毒理学报》2023年第6期217-224,共8页Asian Journal of Ecotoxicology

基　　金：山西省自然科学基金项目(201901D111232);国家自然科学基金项目(31302155)。

摘　　要：砷暴露引起认知障碍的问题已引起科学家的广泛关注。本研究基于转录组测序技术研究砷影响小鼠神经行为学的潜在分子机制。雄性C57BL/6J小鼠随机分为4组:对照组(C组)、低剂量(L组)、中剂量(M组)和高剂量砷暴露组(H组),分别饮含0、0.15、1.5和15 mg·L^(-1)As_(2)O_(3)的去离子水。暴露12周后,采用新事物识别实验评价小鼠认知功能。利用转录组测序获得高剂量组和对照组大脑前额皮质差异表达基因,进行GO分析和KEGG分析,并采用实时荧光定量PCR法验证基因表达水平。构建蛋白互作网络并筛选其中最紧密连接的模块以获得枢纽基因。结果显示,砷暴露组小鼠探索指数和对新旧事物的分辨指数与对照组相比显著降低。共筛选出181个差异表达基因,GO分析得到包括行为学、多细胞生物过程的调节、学习或记忆、认知等217条显著富集条目。KEGG分析获得包括胆碱能突触、多巴胺突触、神经营养素信号通路等38条通路。筛选蛋白互作网络中最紧密连接的模块共获得7个枢纽基因,分别为fos、tac1、gad2、drd2、ppp1r1b、chat和drd1a。实时荧光定量PCR验证了测序结果的可靠性。砷暴露显著降低了小鼠的认知功能。本研究筛选出的关键基因和通路为深入研究砷影响小鼠神经行为学改变的机制提供了线索。Arsenic exposure-induced cognitive disorders have attracted much attention from scientists.This study employed RNA sequencing to investigate the potential molecular mechanisms underlying neurobehavioral alterations in mice due to arsenic exposure at a transcriptomic level.Male C57BL/6 mice were divided into four groups,the control group(group C),low-dose group(group L),medium-dose group(group M)and high-dose group(group H),which drank deionized water containing 0,0.15,1.5 and 15 mg·L^(-1)As_(2)O_(3)respectively for 12 weeks.Cognitive function of the mice was analyzed by the novel object recognition test.RNA sequencing followed by GO enrichment analysis and KEGG pathway enrichment analysis identified differentially expressed genes(DEGs)between control and high dose groups.qRT-PCR was conducted to verify the gene expression levels of target genes.A protein-protein interaction network(PPI)was constructed to identify hub genes from the gene module with the highest connection degree.The findings indicated a significant decrease in both exploration and discrimination indices for arsenic-exposed groups compared to controls.There were 181 DEGs in the high-dose group compared with the control.A total of 217 significantly enriched GO terms were obtained including behavior,regulation of multicellular biological processes,learning and memory,cognition,etc.KEGG enrichment analysis showed that 128 DEGs were enriched in 38 pathways,including cholinergic synapses,dopamine synapses,neurotrophic factor signaling pathways,etc.7 hub genes were screened from the gene module with the highest connection degree,which were fos,tac1,gad2,drd2,ppp1r1b,chat and drd1a.The results of gene expression levels from qRT-PCR are in good agreement with those from RNA-seq,reflecting the reliability of RNA-seq.It was observed that arsenic exposure led to a significant reduction in mice’s cognitive abilities.The identified hub genes and key pathways from this study offer valuable insights into exploring the mechanisms underlying neurobehavioral chan

关 键 词：砷 小鼠 神经行为学 前额皮质 转录组 

分 类 号：X171.5[环境科学与工程—环境科学]