Differential Expression And Preliminary Analysis Of Gene Of Apoptosis Chondrocytes Induced By Interleukin-1beta (IL-1β) Inhibited By Oligomeric Proanthocyanidins

Objective:Using high-throughput sequencing technology to screen differentially expressed m RNA of apoptotic chondrocytes induced by OPC(Oligomeric Proanthocyanidins)inhibited IL-1?(interleukin-1beta,interleukin-1beta)and useing bioinformatics analysis to reveal the possible key regulatory pathways of oligomeric proanthocyanidins in inhibiting the development of chondrocytes,and to explore the role of oligomeric proanthocyanidins in protecting chondrocytes.Methods:1.There are six samples contained in the GSE104793 data Consists of three untreated primary mouse articular chondrocytes and three articular chondrocytes with IL-1β,gene differential expression analysis and KEGG enrichment analysis were performed on the two sets of data.2.Extract and identify primary mouse cartilage cells.3.The control group were added with IL-1β,the experimental group was treated with IL-1β and OPC for 24 hours,extracting m RNA and both of groups were subjected to high-throughput sequencing.4.Screen differentially expressed genes between the experimental group and the control group,perform GO enrichment analysis and KEGG signal pathway enrichment analysis on differentially expressed m RNAs,screen for important differentially expressed m RNAs and regulatory pathways.5.Compare with differentially expressed genes,and screen for genes which reduced expression in GSE104793 and increased expression in sequencing data,or genes which increased expression in GSE104793 and decreased expression in sequencing data;perform GO enrichment analysis about the above genes.Results:1.Analysis of the m RNA expression of GSE104793 data revealed a total of 843 differential m RNAs,497 m RNAs were up-regulated and 346 m RNAs were downregulated in the experimental group compared with the control group.The KEGG signal pathway enrichment analysis of the GSE104793 data found that differentially expressed m RNAs were mainly concentrated in more than Pathways in cancer,Cytokine-cytokine receptor interaction,Cell cycle,Focal adhesion,Cell adhesion molecules(CAMs),ECM-receptor interaction,Toll-like receptor signaling pathway,Chagas disease(American trypanosomiasis),Small cell lung cancer,Progesterone-mediated oocyte maturation,p53 signaling pathway,Rheumatoid arthritis,Melanoma,Bladder cancer,Graft-versus-host disease,DNA replication.2.Immunofluorescence identification of mouse articular chondrocytes showed that the cell purity was greater than 90%.3.The high-throughput sequencing of the experimental group and the control group screened a total of 662 differential m RNAs.Among the experimental group,compared with the control group,256 m RNA expressions were up-regulated and 406 m RNA expressions were down-regulated.All sequences have a base error rate quality score of more than 30 points,a correct base recognition rate of more than 99.9%,a uniform base content,and good overall reads.4.The GO＿BP signal pathway enrichment analysis of the original data found that the biological process were mainly concentrated on more than 10 signal pathways as leukocyte migration,regulation of ERK1 and ERK2 cascade,ERK1 and ERK2 cascade,response to virus,positive regulation of response to external stimulus,viral process,defense response to virus,,negative regulation of cytokine production,myeloid leukocyte migration,mononuclear cell migration.The GO＿CC signal pathway enrichment analysis of raw data were mainly involved in the role of cell components of extracellular matrix,membrane raft,membrane microdomain,membrane region,collagen-containing extracellular matrix,Golgi cisterna,Golgi stack,endoplasmic reticulum exit site,MHC class I protein complex,MHC protein complex.The KEGG signal pathway enrichment analysis of the original data found that differentially expressed m RNAs were mainly concentrated in more than 10 signal pathways as Herpes simplex virus 1 infection,Cytokine-cytokine receptor interaction,Epstein-Barr virus infection,Focal adhesion,Viral protein interaction with cytokine and cytokine receptor,NF-kappa B signaling pathway,Rheumatoid arthritis,Viral myocarditis,Allograft rejection,Graft-versus-host disease.5.The genes that were compared and screened with GSE104793 were: Fjx1、Igfbp5、Chac1、Gfod1、Itga2、Acan、Chchd10、Nes、Cspg4、Grpr、Ntrk3、Angpt1、Chl1、Igf1、Il18rap、Eda2r、Lrch2、Pappa、Slitrk6、Adamts5、Fap、Gda、Prss23、Cfh、Ibsp、Kitl、Rspo2、Trp53inp1、Arrdc3、Vgll3、Il1r1、Gpm6b、Stc1、Klhl24、Rnd3、Gas1、Fst、Steap4。.The GO＿BP signal pathway enrichment analysis of genes that were compared with the GSE104793 and screened revealed that the biological processes were mainly concentrated on more than 10 signal pathways asossification,ameboidal-type cell migration,epithelial cell migration,epithelium migration,tissue migration,connective tissue development,bone mineralization,biomineral tissue development,positive chemotaxis,T-helper 1 cell cytokine production.The GO＿CC signal pathway enrichment analysis was performed to analyze the role of cell components mainly in more than four signal pathways of collagen-containing extracellular matrix,insulin-like growth factor ternary complex,insulin-like growth factor binding protein complex,growth factor complex.Conclusion:OPC may mediate multiple targeted genes through the Toll-like receptor / NF-κB signaling pathway and inhibit chondrocyte apoptosis induced by IL-1β.