The Preliminary Screening And Function Analysis Of LncRNA Related With Mouse Peritoneal Fibrosis

Objectives To detect the dynamic expression of lncRNA in normal and peritoneal fibrosis tissue using microarray,and to study the preliminary function of long chain Noncoding RNA with different expression relationship between lncRNA and peritoneal fibrosis.To provide experimental basis for clinical prevention and treatment of peritoneal dialysis related peritoneal fibrosis from the perspective of molecular biology.Methods 30 male C57/BL of 6 weeks were divided into three groups: normal group?Early peritoneal fibrosis group(model group for 4 weeks)?Advanced peritoneal fibrosis group(model group for 6 weeks).Two model groups were injected with 4.5% peritoneal dialysis fluid(Baxter 1ml/10 g body weight).After experimental period detected,each group of mice peritoneal dialysis ultrafiltration volume and drainage liquid protein concentration,calculate the clearance rate of urea and glucose;HE staining and Masson staining observed peritoneal morphological changes;the lncRNAs expression levels were detected by microarray in normal group and model groups separately;finally the differentially expressed lncRNAs were identified by RT-PCR.The data of mRNA were analyzed by GO clustering,by KEGG database for Pathway annotation.Results:(1)Compared with control group,the ultrafiltration of peritoneal dialysis in two peritoneal fibrosis groups were both decreased(p<0.05),Advanced peritoneal fibrosis group compared with early peritoneal fibrosis group,peritoneal dialysis flow decreased significantly(P<0.05)?Compared with control group,two model groups of peritoneal dialysis,drainage of liquid drainage protein concentration increased(P<0.05);Than between the two model groups,drainage of peritoneal dialysis fluid drainage protein concentration is no significant difference.(2)Compared with control group,two peritoneal fibrosis groups of urea,glucose clearance decreased(P < 0.05);Comparison between the two model groups,advanced peritoneal fibrosis,urea,glucose clearance earlier peritoneal fibrosis groups were decreased(P < 0.05).(3)Morphological characteristics of peritoneal tissue were observed under light microscope,Two peritoneal structure in model group were changed,Peritoneal tissue were thickened,Some of the peritoneal mesothelial cells were deformed?mesothelial cell junction were disappeared?fibrous tissue were proliferative,Interstitial inflammatory cell infiltrated,and with the progress of the peritoneal fibrosis,the significant change gradually.(4)The expressions of lncRNAs and mRNAs were tested by Microarray.The expression of IncRNAs in 2 model groups of peritoneal fibrosis mice were compared with normal group.Two model groups compared with normal group,the results which were the same as the change of lncRNAs were selected.The 4 weeks model group was compared with the control group: a total of 857 lncRNAs were differentially expressed more than 2 times.Among them,341 lncRNAs were up-regulated,516 lncRNAs were down regulated.The 6 weeks model group was compared with the control group: a total of 1071 lncRNAs were differentially expressed more than 2 times,493 lncRNAs were up-regulated,578 lncRNAs were down-regulated.The 6 weeks model group was compared with the 4 weeks group: total of 715 lncRNAs were differentially expressed more than 2 times: 301 lncRNAs were up-regulated and 414 lncRNAs were down-regulated.Compared with the control group,there were 1047 mRNA with different expression more than 2 times in 4weeks model group,279 mRNAs were up-regulated,768 mRNAs were down regulat-ed.Compare the 6 weeks model group with the control group,total of 1411 mRNAs were differentially expressed more than 2 times,502 mRNAs were up-regulated,809 mRNAs were down-regulated.The 6 weeks model group was compared with the 4 weeks group: total of 689 mRNAs were differentially expressed more than 2 times,165 mRNAs were up-regulated and 524 mRNAs were down-regulated.(5)GO cluster analysis results show: In the process of peritoneal dialysis correlation peritoneal fibrosis,differentially expressed genes were mainly involved in the cell membrane of the inherent composition,the composition of cells surrounding the organizational structure,the inflammation of the defense reaction,ion transport,etc.Query the different significant genes in KEGG database,Set P < 0.05 for significant standard,differentially expressed genes may be involved in eukaryotes translation process,the way such as membrane protein synthesis.(6)RT-PCR verified differentially expressed genesNONMMUT024530?NONMMU T002955?NONMMUT055611,The trend was consistent with the chip results Conclusion:In the expression of peritoneal fibrosis in mice tissues,lncRNAs spectru-um dynamic changes have taken place significantIly,Prompt lncRNAs may participate in the occurrence and development of peritoneal fibrosis gene regulation.