| Background and purpose:Abdominal adhesion is a common postoperative problem in surgery.Abdominal adhesion in children may cause serious complications such as adhesive intestinal obstruction,intestinal strangulation and intestinal necrosis.Especially female children may cause adhesion of pelvic organs and affect long-term fertility.The treatment of complications caused by abdominal adhesion has some problems,such as the poor effect of conservative treatment,large trauma of surgical treatment and repeated intestinal obstruction after operation,which is not conducive to children’s growth and mental health.At present,the research on the mechanism of abdominal adhesion is still under continuous improvement and development.With the openness and transparency of information,a large number of public databases have been gradually improved,and a large number of scholars have used bioinformatics to mine disease information around the world.The purpose of this study is to screen the key genes for the formation of abdominal adhesion through bioinformatics technology,verify its expression level with clinical samples,and explore the regulatory relationship between Fn1(Fibronectin 1,Fnl),and PI3K(Phosphatidylinositol3-kinase,PI3K)/Akt(Protein Kinase B,Akt/PKB)at the cellular level,so as to provide new ideas for targeted prevention and treatment of abdominal adhesion in children.Methods:1.Download GSE123413 data set from GEO database,and use R language to perform differential expression analysis,weighted gene co-expression network analysis and Wayne analysis to screen the core genes of abdominal adhesion;GO enrichment analysis and KEGG signal pathway analysis of core genes were carried out by R language.Finally,the protein Interaction Network(Protein-Protein Interaction Networks,PPI)is constructed with the STRING database.The results were visualized by Cytoscape software and the top 10 proteins were screened out.2.HE staining and Masson staining were used to treat the tissue of abdominal adhesions and analyze its components.3.Quantitative reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry were used to detect the mRNA level of Fnl in clinical samples of peritoneal adhesion and normal peritoneum.4.The human peritoneal mesothelial cells were down-regulated by cell transfection,and then the expressions of Fnl,PI3K,Akt,p-PI3K and p-Akt were analyzed by quantitative reverse transcription polymerase chain reaction and protein blot.Results:1.A total of 368 common module genes closely related to the phenotype of celiac adhesion were screened from GSE123413 data set,and finally 10 core genes of celiac adhesion(Fnl,Col1a1,Col1a2,Col3a1,Acta2,Pcolce,Serpinhl,Col2a1,Dcn and Fbn1)were obtained,among which Fnl had the highest correlation.2.By HE and Masson staining,the composition of the adhesive ligament in the abdominal cavity was analyzed.The results showed that the peritoneal tissue was continuously destroyed,thickened in different degrees,and the fibrous tissue was dense,and the collagen fiber components in it were obviously deposited and stained deeply.Infiltration of inflammatory cells and new blood vessels can be seen in the tissue.3.Quantitative reverse transcription polymerase chain reaction and immunohistochemical method showed that Fnl expression was significantly upregulated in abdominal adhesion tissue.4.In human peritoneal mesothelial cells,after down-regulating Fnl by transfection,the expression levels of PI3K and Akt did not change significantly,but the phosphorylated PI3K and Akt decreased significantly,which was statistically significant.Conclusions:1.Ten core genes of abdominal adhesion were screened,among which Fnl had the highest correlation.2.The expression level of Fnl in peritoneal adhesion tissue was high.3.During the formation of abdominal adhesion,Fnl may promote the function of mesothelial cells by influencing the phosphorylation of PI3K and Akt,which lays a foundation for future targeted prevention of abdominal adhesion. |
PDF Full Text Request