To Screen Differentially-expressed Genes Of Genipin On Hippocampus And Frontal Cortex In Mice Based On RNA-seq Technology

Background:Genipin is present in many traditional Chinese medicines and is the main active ingredient in gardenia.The earliest studies have shown that it is a good biocrosslinker and can be used as a biomaterial in clinical applications.With the continuous deepening of the research on Genipin,its pharmacological effects such as protecting the liver,fighting inflammation,protecting the stomach,lowering blood sugar,gallbladder,inhibiting neurotoxicity,and suppressing depression have been gradually confirmed.However,the molecular changes related to the antidepressant effect of genipin have not been mentioned,so studying the molecular changes of anti-depression of genipin further contributes to its clinical application as an antidepressant.Depression as one of the affective disorder diseases,its main clinical manifestations are significant and lasting depressed mood,and some seriously ill patients have suicidal tendency and seriously endanger human health.According to 2017 reports from World Health Organization,the incidence of depression is gradually increasing,with about 350 million people.In the comprehensive ranking of disability and death,depression is ranked after heart disease,Stroke and AIDS.At present,the etiology andpathogenesis of depression and the treatment of antidepressants have become a research hotspot.RNA-seq is a high-throughput sequencing technology currently widely used in the study of transcriptomics.Its main functions include: detection of unknown genes,quantification of gene expression levels,discovery of new transcripts,and accurate identification of variable cleavage sites.Points and cSNP,UTR area.Functional detection of differential gene expression screening is currently used most.Therefore,RNA-seq has a good application prospect in clinical research work.Objective: To screendiffrentially-expressed genes of genipin on hippocampus and forebrain cortex in mice by RNA-seq technology,and to provide basis for further investigation of antidepressant effect of genipin.Method:1.Male Kunming mice of 4 weeks of age were selected as experimental subjects.According to the principle of random grouping,mice were divided into genipin group and control group.Experimental group: genipin ip,control group: DMSO ip,daily 1 time,7 days in a row.2.RNA-seq technology was used to screen the differential gene expression of genipin in the hippocampus and forebrain cortex of KM mice and control groups.3.Real-time quantitative PCR technique was used to verify the reliability of the differentially expressed genes with significant significance in sequencing,and to understand the possible related genes and related pathways of genipin.Result:1.There were 151 and 524 genes differentially expressed in the hippocampus and forebrain cortex between the two groups after genipintreatment,and there were 42 differential genes overlapping in the two tissues.The differentially expressed genes in the forebrain of the genipin-treated group contained 248 genes up-regulated and 276 genes were down-regulated.At the same time,91 genes were differentially expressed in the hippocampus,and 60 genes were down-regulated.Possible related pathways include intracellular Ca signaling,neural spine development,regulation of biological rhythms,and depression-related pathways.2.Real-time fluorescence quantitative PCR validation of differentially expressed genes,including: Mobp,Sirt2,Per3,Itgad,Hdac1,Fkbp10,Htr2 a,Egr2,Egr3,Egr4,Nos1,Grin2 c,Grm3,Grm5,Cacnb1,Hist2h2 bb,Hist2h3c2,Hist1h2 bc,Miat,the above gene change trend is basically the same as the sequencing result.Conclusion:There were 151 and 524 differentially expressed genes in the mouse hippocampus and forebrain cortex after genipin treatment.There were 42 differential genes that overlapped in two tissues and were included in depression-related genes and pathways.