RESUMEN
Uridine diphosphate glycosyltransferase(UGT) is involved in the glycosylation of a variety of secondary metabolites in plants and plays an important role in plant growth and development and regulation of secondary metabolism. Based on the genome of a diploid Chrysanthemum indicum, the UGT gene family from Ch. indicum was identified by bioinformatics methods, and the physical and chemical properties, subcellular localization prediction, conserved motif, phylogeny, chromosome location, gene structure, and gene replication events of UGT protein were analyzed. Transcriptome and real-time fluorescence quantitative polymerase chain reaction(PCR) were used to analyze the expression pattern of the UGT gene in flowers and leaves of Ch. indicum. Quasi-targeted metabolomics was used to analyze the differential metabolites in flowers and leaves. The results showed that a total of 279 UGT genes were identified in the Ch. indicum genome. Phylogenetic analysis showed that these UGT genes were divided into 8 subfamilies. Members of the same subfamily were distributed in clusters on the chromosomes. Tandem duplications were the main driver of the expansion of the UGT gene family from Ch. indicum. Structural domain analysis showed that 262 UGT genes had complete plant secondary metabolism signal sequences(PSPG box). The analysis of cis-acting elements indicated that light-responsive elements were the most ubiquitous elements in the promoter regions of UGT gene family members. Quasi-targeted metabolome analysis of floral and leaf tissue revealed that most of the flavonoid metabolites, including luteolin-7-O-glucoside and kaempferol-7-O-glucoside, had higher accumulation in flowers. Comparative transcriptome analysis of flower and leaf tissue showed that there were 72 differentially expressed UGT genes, of which 29 genes were up-regulated in flowers, and 43 genes were up-regulated in leaves. Correlation network and phylogenetic analysis showed that CindChr9G00614970.1, CindChr2G00092510.1, and CindChr2G00092490.1 may be involved in the synthesis of 7-O-flavonoid glycosides in Ch. indicum, and real-time fluorescence quantitative PCR analysis further confirmed the reliability of transcriptome data. The results of this study are helpful to understand the function of the UGT gene family from Ch. indicum and provide data reference and theoretical basis for further study on the molecular regulation mechanism of flavonoid glycosides synthesis in Ch. indicum.
Asunto(s)
Chrysanthemum , Glicosiltransferasas , Glicosiltransferasas/genética , Glicosiltransferasas/metabolismo , Chrysanthemum/genética , Uridina Difosfato , Filogenia , Reproducibilidad de los Resultados , Plantas/metabolismo , Flavonoides , Glicósidos , Regulación de la Expresión Génica de las PlantasRESUMEN
Based on the record of Miu Ci theory from Huangdi Neijing (The Yellow Emperor's Internal Classic), and incorporating the relevant discussions of medical scholars from various dynasties, this article interprets and analyzes the Miu Ci technique, the points to be needled, and the diseases can be treated. The following innovative understandings are proposedï¼ 1) The original meaning of "Miu" in Miu Ci is "to prick in a different way from the meridians (needle the major collaterals)", not "needle left and right interchangeably". Needle left and right interchangeably is not a necessary operation in Miu Ci. 2) The stimulation sites of Miu Ci are the "four extremities" of the human body, referred to as the "major collaterals of qi", and the nail bed of the four extremities is not equivalent to the commonly reco-gnized "Jing-well points". 3) Miu Ci can treat critical illnesses, not just limited to musculoskeletal pain or mild conditions at the early stage of a disease. 4) Miu Ci is not equivalent to Luo Ci needling (pricking bloodletting therapy).
Asunto(s)
Terapia por Acupuntura , Meridianos , Humanos , Medicina Tradicional China , Terapia por Acupuntura/métodos , Venodisección , Agujas , Puntos de AcupunturaRESUMEN
N6-methyladenosine (m6A) methylation is a vital epigenetic mechanism associated with drug addiction. However, the relationship between m6A modification and oxycodone rewarding is less well explored. Based on an open field test, the present study evaluated oxycodone rewarding using chromatin immunoprecipitation PCR, immunofluorescence, and RNA sequencing. A marked increase in METTL14 protein and a decrease in PP1α protein due to oxycodone abundance in the striatal neurons were observed in a dose- and time-dependent manner. Oxycodone markedly increased LSD1 expression, and decreased H3K4me1 expression in the striatum. In the open field test, intra-striatal injection of METTL14 siRNA, HOTAIR siRNA, or LSD1 shRNA blocked oxycodone-induced increase in locomotor activity. The downregulation of PP1α was also inhibited after treatment with METTL14/HOTAIR siRNA and LSD1 shRNA. Enhanced binding of LSD1 with CoRest and of CoRest with the PP1α gene induced by oxycodone was also reversed by LSD1 shRNA. In addition, H3K4me1 demethylation was also blocked by the treatment. In summary, the investigation confirmed that METTL14-mediated upregulation of HOTAIR resulted in the repression of PP1α, which in turn facilitated the recruitment of LSD1, thus catalyzing H3K4me1 demethylation and promoting oxycodone addiction.
Asunto(s)
Metiltransferasas , Oxicodona , ARN Largo no Codificante , Animales , Masculino , Ratones , Cuerpo Estriado/metabolismo , Cuerpo Estriado/efectos de los fármacos , Desmetilación , Histona Demetilasas/metabolismo , Histona Demetilasas/genética , Histonas/metabolismo , Lisina/análogos & derivados , Metiltransferasas/metabolismo , Metiltransferasas/genética , Ratones Endogámicos C57BL , Oxicodona/farmacología , Proteína Fosfatasa 1/metabolismo , Proteína Fosfatasa 1/genética , ARN Largo no Codificante/metabolismo , ARN Largo no Codificante/genética , Regulación hacia ArribaRESUMEN
The lysine-specific demethylase 1 (KDM1A) is reported to be a regulator in learning and memory. However, the effect of KDM1A in oxycodone rewarding memory has yet to be studied. In our study, rewarding memory was assessed by using conditioned place preference (CPP) in male mice. Next generation sequencing and chromatin immunoprecipitation-PCR were used to explore the molecular mechanisms. Oxycodone significantly decreased PP1α mRNA and protein levels in hippocampal neurons. Oxycodone significantly increased KDM1A and H3K4me1 levels, while significantly decreased H3K4me2 levels in a time- and dose-dependent manner. Behavioral data demonstrated that intraperitoneal injection of ORY-1001 (KDM1A inhibitor) or intra-hippocampal injection of KDM1A siRNA/shRNA blocked the acquisition and expression of oxycodone CPP and facilitated the extinction of oxycodone CPP. The decrease of PP1α was markedly blocked by the injection of ORY-1001 or KDM1A siRNA/shRNA. Oxycodone-induced enhanced binding of CoRest with KDM1A and binding of CoRest with the PP1α promoter was blocked by ORY-1001. The level of H3K4me2 demethylation was also decreased by the treatment. The results suggest that oxycodone-induced upregulation of KDM1A via demethylation of H3K4me2 promotes the binding of CoRest with the PP1α promoter, and the subsequent decrease in PP1α expression in hippocampal neurons may contribute to oxycodone reward.