海洛因诱导条件性位置偏爱大鼠伏隔核的转录组学研究

Transcriptomics study of heroin-induced conditioned place preference in the nucleus accumbens of rats

目的研究海洛因依赖对大鼠伏隔核的转录组和代谢组的影响。方法将8只SD大鼠随机分成生理盐水对照组和海洛因诱导组,两组大鼠均采用剂量递增法分别经皮下注射生理盐水和海洛因,连续注射7 d,建立海洛因诱导位置偏爱模型。实验第8天将2组大鼠断头取脑,剥离伏隔核,提取RNA进行转录组学检测,分析海洛因依赖大鼠伏隔核相关基因的差异性表达。结果与对照组比较,根据DESeq2 P值<0.05 log2样本间表达量比值>0筛选标准,海洛因依赖大鼠的测序数据共926个差异基因,其中上调基因410个,下调基因516个。GO功能富集中,生物过程(BP)分析,与对照组相比,Heroin组在突触可塑性的调节、谷氨酸受体信号通路的调节、长时程突触增强效力等方面上调,在固醇、酒精及有机羟基化合物等的代谢及生物合成过程方面下调。细胞组成(CC)方面提示谷胱甘肽突触基因表达升高,核糖体、内线粒体膜蛋白复合物等基因表达下降。分子功能(MF)中,糖皮质激素受体结合基本生化活性中的基因表达升高,核糖体的结构成分等基因降低。KEGG通路富集中海洛因组在MAPK信号通路、黑色素生成和胆碱能突触等方面上调;在类固醇生物合成、核糖体、剪接体、缬氨酸、亮氨酸和异亮氨酸降解以及MAPK信号通路等方面下调。结论突触可塑性和谷氨酸受体信号通路的调节、长时程增强及MAPK信号通路在海洛因诱导CPP状态大鼠伏隔核中发挥重要作用。

Objective To study the effects of heroin addiction on the transcriptome and metabolome of the rat nucleus accumbens.Methods Eight SD rats were randomly divided into two groups,namely the saline control group and the heroin induction group.Both groups of rats were injected with saline and heroin subcutaneously by increasing doses for seven consecutive days to establish a heroin-induced conditioned place preference(CPP)model.On the 8th day of the experiment,the two groups of rats were decapitated and their brains were stripped of the nucleus accumbens,RNA was extracted for transcriptomics testing,and the differential expression of genes related to the nucleus accumbens of heroin-dependent rats was analyzed.Results Compared with the control group,according to the screening criteria of DESeq2 P-value<0.05 log2FoldChange>0,there were a total of 926 differential genes in the sequencing data of heroin-dependent rats,of which 410 were up-regulated genes and 516 were down-regulated genes.Through GO functions enrichment and biological process(BP)analysis,compared with the control group,the Heroin group is up-regulated in the regulation of synaptic plasticity,the regulation of glutamate receptor signaling pathways,and the potency of long-term synaptic enhancement.The metabolism and biosynthesis processes of alcohol,alcohol and organic hydroxy compounds are down-regulated.Cell composition(CC)suggests that the expression of glutathione synaptic genes is increased,and the expressions of genes such as ribosomes and inner mitochondrial membrane protein complexes are decreased.In molecular function(MF),the expressions of genes involved in glucocorticoid receptor binding to basic biochemical activity are increased,and genes such as the structural components of ribosomes are decreased.KEGG pathway enriched heroin group is up-regulated in MAPK signaling pathway,melanogenesis,cholinergic synapse,etc.;in steroid biosynthesis,ribosome,spliceosome,valine,leucine and isoleucine degradation,and MAPK signaling Decrease in access and other aspects.Conclusion Synaptic plasticity and regulation of glutamate receptor signaling,longterm synaptic potentiation,and MAPK signaling play important roles in the nucleus accumbens of heroin-induced CPP rats.