Analysis Of MiRNAs Expression Profile And Prediction Of Target Signaling Pathway In Islet Cells Of Rat Pancreatic Head And Pancreatic Tail

Research of the differential expression of miRNAs in the pancreatic head and pancreatic islet cells of the rat and study the prediction of target genes and analysis of signaling pathways.In order to discuss the further exploration of the role of miRNAs physiology and pathology in islet.Methods1.Randomly select 6 normal adult rats from 6 to 24 weeks,take the head of the pancreas and the tail of the pancreas,separate the islets by density gradient centrifugation,pick up the islet cells by hand and confirm by microscopic examination;2.Send islet cells to Shanghai Kangcheng Biological Company for high-throughput sequencing analysis of miRNA to detect the expression level of miRNAs;combined with bioinformatics methods to screen out significantly differentially expressed miRNAs and target differentially expressed miRNAs through Targetscan and miRDB Gene prediction,using the VENNY online database to obtain the target genes supported by the intersection of the two genes predicted by the two databases,and performing functional analysis of the GO and KEGG databases;enrichment analysis of the target gene proteins through the DAVID database,Screen out miRNAs with statistical differences and clinical significance.3.Real-time fluorescence quantitative PCR(RT-PCR)verification of the selected miRNA,and analysis of its potential physiological and clinical significance to the pancreatic head and the pancreatic tail by target genes and pathways.Result1.A total of 445 miRNAs with statistical significance were detected by miRNA sequencing.Using the pancreatic tail as a control group,a total of 376 non-significant miRNAs were screened out,and 69 miRNAs were significantly differentially expressed(of which 28 were up-regulated,There are 41 reductions).2.The results of GO enrichment analysis showed that the target gene enrichment regulated by differential expression of miRNAs at the head and end of the pancreas differed in terms of positioning,molecular function,and biological process.From the KEGG enrichment analysis,it can be seen that the highest enrichment of the pancreatic head-end pathway is the FOXO signaling pathway,in which 26 differential target genes are enriched,which shows that the FOXO signaling pathway may be the most statistically significant for the pancreatic head and pancreatic tail Differential pathways of meaning.3.According to data statistics and sequencing analysis,there are statistically significant differences in the miR-124 at the pancreatic head and pancreatic end,and the content of miR-124 at the pancreatic head is significantly higher than that of the pancreas.In view of the miR-124 enriched pathways and target genes,and their important role in the regulation of pancreatic physiology and pancreatic cancer,it shows that it is enriched in the insulin secretion signaling pathway by regulating the target genes ATP1A1,ITPR3,and RYR;Gene Flotillin-1,TC10,AMPK,SOS,PP1,PHK,SHC,etc.are enriched in the insulin signaling pathway,and through target genes NF-κB,STAT3,ECAD,ITGB,Rhn GEF,VEGF,ECM,AR,RAl BP1,TRAFs,etc Enriched in cancer signal pathway;real-time fluorescent quantitative PCR verification results indicate that the expression level of miR-124 at the pancreatic head and pancreatic tail is statistically different,which is consistent with the sequencing results.Conclusion1.In addition to the differences in the anatomical structure and physiological function of pancreatic head and the pancreatic tail,the sequencing results show that there are also significant statistical differences in the expression of miRNAs between pancreatic head and the pancreatic tail.2.The results of GO and KEGG enrichment analysis suggest that among the target gene enrichment pathway,the FOXO signaling pathway is highly expressed in the head of the pancreas than the tail of the pancreas.3.According to the sequencing results and RT-PCR verification tips,there are statistical differences in the expression of miR-124 at the pancreatic head and pancreatic tail,and the expression of miR-124 at the pancreatic head is significantly higher than that of the pancreas.