目的采用代谢组学方法研究丁卡因急性中毒致死小鼠血清和组织(肾、肝和心脏)中的代谢物变化,寻找潜在的生物标志物及其相关代谢通路,为丁卡因急性中毒的死亡原因鉴定及毒理机制研究提供新思路。方法40只ICR小鼠被随机分为对照组和丁卡...目的采用代谢组学方法研究丁卡因急性中毒致死小鼠血清和组织(肾、肝和心脏)中的代谢物变化,寻找潜在的生物标志物及其相关代谢通路,为丁卡因急性中毒的死亡原因鉴定及毒理机制研究提供新思路。方法40只ICR小鼠被随机分为对照组和丁卡因急性中毒致死组,以腹腔注射丁卡因建立急性中毒致死模型,运用超高效液相色谱-静电场轨道阱高分辨质谱联用(ultra-high performance liquid chroma⁃tography-electrostatic field orbitrap high resolution mass spectrometry,UPLC-Orbitrap HRMS)获取小鼠血清和组织的代谢轮廓。利用多元变量统计主成分分析和正交偏最小二乘-判别分析,并结合t检验和差异倍数分析找出与丁卡因急性中毒致死相关的差异代谢物。结果与对照组相比,丁卡因急性中毒致死组小鼠血清和组织的代谢轮廓表现出明显的区分。血清中鉴定出11种差异代谢物,包括黄嘌呤、精胺、3-羟基丁胺等;肝组织中鉴定出25种差异代谢物,包括腺苷酸、腺苷、柠檬酸等;心脏中鉴定出12种差异代谢物,包括次黄嘌呤、鸟嘌呤、鸟苷酸等;肾组织中鉴定出4种差异代谢物,包括牛磺鹅去氧胆酸、11,12-环氧二十碳三烯酸、二甲基乙醇胺和吲哚。丁卡因急性中毒主要影响了嘌呤代谢,三羧酸循环,丙氨酸、天冬氨酸和谷氨酸代谢。结论丁卡因急性中毒致死小鼠血清和组织中的差异代谢物有望成为该死因的候选生物标志物,该结果可为丁卡因急性中毒的作用机制和死亡原因鉴定提供研究基础。展开更多
OBJECTIVE Cue-induced drug craving progressively increase after prolong abstinence in animal and human beings.A behavioral phenomenon termed incubation of drug craving is considered as a key reason for drug relapse,bu...OBJECTIVE Cue-induced drug craving progressively increase after prolong abstinence in animal and human beings.A behavioral phenomenon termed incubation of drug craving is considered as a key reason for drug relapse,but the transcriptional mechanisms that contribute to this incubation are unknown.It has been demonstrated that circular RNAs(circR NAs),act as miR NA sponges,play important roles in the regulation of gene expression and the pathogenesis of disease.The present study aims to explore the transcriptional profiles associated with incubation of morphine craving in nucleus accumbens(NAc),an important brain area previously implicated in drug seeking.METHODS The animal model of the incubation of drug craving was induced by CPP paradigms with six morphine(5 mg·kg-1) injections and 14 d drug abstinence in mice.The brain tissues of NAc were collected after the behavioral tests for circRNA-sequencing by Illumina Hiseq X sequencer.We identified differentially expressed genes(DEGs) by qRT-PCR and used bioinformatics methods for further function analysis.RESULTS The progressive increase of CPP scores during the 14 d drug abstinence indicated the establishment of animal model.The data of circRNA-sequencing reported that 16 circRNAs were significantly altered after 28 d drug abstinence in NAc of morphine treated mice(FC≥2 and P<0.05).Among those circRNAs,9 were significantly up-regulated,while 7 were down-regulated.Furthermore,we subsequently tested circRNAs expression using quantitative real-time PCR,and the consistent data were obtained.The results of KEGG pathway analysis indicated that these genes were enriched for several biological processes,including RNA transport,protein ubiquitination and histone methylation,etc.CONCLUSION These findings provide a unique resource of gene expression data for future studies examining transcriptional mechanisms in NAc that mediate opioids seeking after prolonged withdrawal.展开更多
Background It is known that excessive release of glutamate can induce excitotoxicity in neurons and lead to seizure. Dexamethasone has anti-seizure function. The aim of this study was to investigate glutamatedexametha...Background It is known that excessive release of glutamate can induce excitotoxicity in neurons and lead to seizure. Dexamethasone has anti-seizure function. The aim of this study was to investigate glutamatedexamethasone interaction in the pathogenesis of epilepsy, identify differentially expressed genes in the hippocampus of glutamate-induced epileptic rats by mRNA differential display, and observe the effects of dexamethasone on these genes expression. Methods Seizure models were established by injecting 5μl （250 μg/μl） monosodium glutamate （MSG） into the lateral cerebral ventricle in rats. Dexamethasone （5 mg/kg） was injected intraperitoneally at 30 minutes after MSG inducing convulsion. The rats＇ behavior and electroencephalogram （EEG） were then recorded for 1 hour. The effects of dexamethasone on gene expression were observed in MSG-induced epileptic rats at 1 hour and 6 hours after the onset of seizure by mRNA differential display. The differentially expressed genes were confirmed by Dot blot. Results EEG and behaviors showed that MSG did induce seizure, and dexamethasone could clearly alleviate the symptom, mRNA differential display showed that MSG increased the expression of some genes in epileptic rats and dexamethasone could downregulate their expression. From more than 10 differentially expressed eDNA fragments, we identified a 226 bp eDNA fragment that was expressed higher in the hippocampus of epileptic rats than that in the control group. Its expression was reduced after the administration of dexamethasone. Sequence analysis and protein alignment showed that the predicted amino acid sequence of this cDNA fragment kept 43% identity to agmatinase, a member of the ureohydrolase superfamily. Conclusions The results of the current study suggest that the product of the 226 bp eDNA has a function similar to agmatinase. Dexamethasone might relax alleviate seizure by inhibiting expression of the gene.展开更多
文摘目的采用代谢组学方法研究丁卡因急性中毒致死小鼠血清和组织(肾、肝和心脏)中的代谢物变化,寻找潜在的生物标志物及其相关代谢通路,为丁卡因急性中毒的死亡原因鉴定及毒理机制研究提供新思路。方法40只ICR小鼠被随机分为对照组和丁卡因急性中毒致死组,以腹腔注射丁卡因建立急性中毒致死模型,运用超高效液相色谱-静电场轨道阱高分辨质谱联用(ultra-high performance liquid chroma⁃tography-electrostatic field orbitrap high resolution mass spectrometry,UPLC-Orbitrap HRMS)获取小鼠血清和组织的代谢轮廓。利用多元变量统计主成分分析和正交偏最小二乘-判别分析,并结合t检验和差异倍数分析找出与丁卡因急性中毒致死相关的差异代谢物。结果与对照组相比,丁卡因急性中毒致死组小鼠血清和组织的代谢轮廓表现出明显的区分。血清中鉴定出11种差异代谢物,包括黄嘌呤、精胺、3-羟基丁胺等;肝组织中鉴定出25种差异代谢物,包括腺苷酸、腺苷、柠檬酸等;心脏中鉴定出12种差异代谢物,包括次黄嘌呤、鸟嘌呤、鸟苷酸等;肾组织中鉴定出4种差异代谢物,包括牛磺鹅去氧胆酸、11,12-环氧二十碳三烯酸、二甲基乙醇胺和吲哚。丁卡因急性中毒主要影响了嘌呤代谢,三羧酸循环,丙氨酸、天冬氨酸和谷氨酸代谢。结论丁卡因急性中毒致死小鼠血清和组织中的差异代谢物有望成为该死因的候选生物标志物,该结果可为丁卡因急性中毒的作用机制和死亡原因鉴定提供研究基础。
基金Natural Science Foundation of Hebei Province(H2018206166)National Natural Science Foundation of China (81871524).
文摘OBJECTIVE Cue-induced drug craving progressively increase after prolong abstinence in animal and human beings.A behavioral phenomenon termed incubation of drug craving is considered as a key reason for drug relapse,but the transcriptional mechanisms that contribute to this incubation are unknown.It has been demonstrated that circular RNAs(circR NAs),act as miR NA sponges,play important roles in the regulation of gene expression and the pathogenesis of disease.The present study aims to explore the transcriptional profiles associated with incubation of morphine craving in nucleus accumbens(NAc),an important brain area previously implicated in drug seeking.METHODS The animal model of the incubation of drug craving was induced by CPP paradigms with six morphine(5 mg·kg-1) injections and 14 d drug abstinence in mice.The brain tissues of NAc were collected after the behavioral tests for circRNA-sequencing by Illumina Hiseq X sequencer.We identified differentially expressed genes(DEGs) by qRT-PCR and used bioinformatics methods for further function analysis.RESULTS The progressive increase of CPP scores during the 14 d drug abstinence indicated the establishment of animal model.The data of circRNA-sequencing reported that 16 circRNAs were significantly altered after 28 d drug abstinence in NAc of morphine treated mice(FC≥2 and P<0.05).Among those circRNAs,9 were significantly up-regulated,while 7 were down-regulated.Furthermore,we subsequently tested circRNAs expression using quantitative real-time PCR,and the consistent data were obtained.The results of KEGG pathway analysis indicated that these genes were enriched for several biological processes,including RNA transport,protein ubiquitination and histone methylation,etc.CONCLUSION These findings provide a unique resource of gene expression data for future studies examining transcriptional mechanisms in NAc that mediate opioids seeking after prolonged withdrawal.
基金The study was supported by a grant from the National Natural Science Foundation of China (No. 39330210).
文摘Background It is known that excessive release of glutamate can induce excitotoxicity in neurons and lead to seizure. Dexamethasone has anti-seizure function. The aim of this study was to investigate glutamatedexamethasone interaction in the pathogenesis of epilepsy, identify differentially expressed genes in the hippocampus of glutamate-induced epileptic rats by mRNA differential display, and observe the effects of dexamethasone on these genes expression. Methods Seizure models were established by injecting 5μl （250 μg/μl） monosodium glutamate （MSG） into the lateral cerebral ventricle in rats. Dexamethasone （5 mg/kg） was injected intraperitoneally at 30 minutes after MSG inducing convulsion. The rats＇ behavior and electroencephalogram （EEG） were then recorded for 1 hour. The effects of dexamethasone on gene expression were observed in MSG-induced epileptic rats at 1 hour and 6 hours after the onset of seizure by mRNA differential display. The differentially expressed genes were confirmed by Dot blot. Results EEG and behaviors showed that MSG did induce seizure, and dexamethasone could clearly alleviate the symptom, mRNA differential display showed that MSG increased the expression of some genes in epileptic rats and dexamethasone could downregulate their expression. From more than 10 differentially expressed eDNA fragments, we identified a 226 bp eDNA fragment that was expressed higher in the hippocampus of epileptic rats than that in the control group. Its expression was reduced after the administration of dexamethasone. Sequence analysis and protein alignment showed that the predicted amino acid sequence of this cDNA fragment kept 43% identity to agmatinase, a member of the ureohydrolase superfamily. Conclusions The results of the current study suggest that the product of the 226 bp eDNA has a function similar to agmatinase. Dexamethasone might relax alleviate seizure by inhibiting expression of the gene.
