Preliminary Study On The Heterogeneity Of LncRNA Expression Between Pancreatic Head Islet And Pancreatic Tail Islet On Rats

Objective:1.To investigate whether the Lnc RNA expression in pancreatic head islets and pancreatic tail islets of 24-week-old rats have heterogeneous.2.Use bioinformatics analysis to predict the biological function and its role of differentially expressed Lnc RNA Method:1.Perfuse the Collagenase into the rat pancreatic through the bile duct,density gradient separation after digestion and sieving and hand pick islets to obtain the purified islet cell and use DTZ to identified it.2.Detect the expression of Lnc RNA in islet cells by using Arraystar Rat Lnc RNA Arrays V3.0 Differential expression profiles of Lnc RNA and m RNA in pancreatic islet cells of rat pancreatic head and tail were determined base on the standard that fold change>2 and p<0.05.By reading the relevant literature,we screened out the Lnc RNA that have statistical significance and significant differential expression as the research objectives.3.Through GO analysis and KEGG analysis of differentially expressed m RNA,analyse the biological functions of differential genes.Lnc RNA with significant differential expression obtained by screening was subjected to correlation analysis(pearson correlation coefficient≥0.9)to Predicting co-expressed m RNA and create a co-expression network to predict the molecular function and the possible biological processes of target Lnc RNA.4.Use Real-time fluorescent quantitative PCR(q PCR)to further verify the expression level of target Lnc RNA in islets of 24-week-old rats.Result:1.A total of 6 samples of pancreatic islet cells from the pancreatic head and tail of the 24-week-old rat were successfully obtained after bile duct perfusion,collagenase digestion and density gradient separation.Quality inspection qualified and Send to Shanghai Kangcheng Biological Engineering Company for sequencing.2.Obtain Lnc RNA and m RNA differential expression profile data of rat pancreatic islet and pancreatic tail islet through Arraystar rat Lnc RNA V3.0expression profile microarray.Base on the standard that fold change>2 and p<0.05,compared the pancreatic tail to pancreatic head,there were 163 significantly different Lnc RNAs,of which 99 Lnc RNAs up-regulated,64 Lnc RNAs down-regulated and7829 Lnc RNAs expressions remained unchanged.And There were 605 significantly differentially expressed m RNAs,of which 263 m RNAs were up-regulated,342 m RNAs were down-regulated,and 25858 m RNAs remained unchanged.3.GO and KEGG analysis was used on differentially expressed m RNA.GO analysis showed that compared with the pancreatic head,down-regulated m RNA in pancreatic tail are mainly involved in tissue development,regulating response to stimuli,signal transduction regulation,aging,cell surface receptor signaling pathway,signal receptor binding,protein binding,and cell adhesion molecules Binding,receptor-regulated activity,etc.And the up-regulated m RNA in pancreatic tail is mainly involved in cell surface receptor signaling pathways,regulating ion transmembrane transport,regulation of synaptic signal transductio,regulation of chemical synaptic transmission,oxidoreductase activity,transferase activity,G protein coupled nucleotide receptors Physical activity,RNA polymerase I activity and so on.4.Co-expression analysis of target Lnc RNA and differentially expressed m RNA,with r≥0.90,p<0.05 as standards,A total of 10 m RNAs with the same expression pattern as Lnc RNA uc.471-were screened out;A total of 8 m RNAs with the same expression pattern as Lnc RNA XR＿592246 were screened out.The GO analysis for the co-expressed m RNA revealed that the co-expressed m RNA are mainly involved in the positive regulation of DNA damage response,signal transduction by P53 class mediator and negative regulation of cell cycle5.q PCR results showed that the expression of Lnc RNA uc.471-and Lnc RNA XR＿592246 in the islet tissue are higher in the pancreatic head compaired with pancreatic tai in 24-week-old rat,consistent with the microarray results.Conclusion:1.Differential expression of Lnc RNA and m RNA was found in islet cells of the pancreatic head and tail of the 24-week-old rat.2.Compared with pancreatic tail,Lnc RNA uc.471-and Lnc RNA XR＿592246may be involved in the regulation of pancreatic islet cell apoptosis of pancreatic head.