Identification Of Differentially Expressed MicroRNAs Involved In Trauma-induced Osteonecrosis Of The Femoral Head Through MicroRNA Expression Profiling

Objective: The present study aimed to profile the differential mi RNA expression between TONFH and normal femoral using high-throughput gene chip detection technology and real-time PCR verification and to develop further understanding of the molecular mechanisms involved in the pathogenesis of TONFH.Methods: Peripheral blood plasma was collected from from 5 patients with TIONFH and 5 with healthy volunteers.The RNA extracted from plasma used to analyze the mi RNA expression profiles using the Exiqon mi RCURY? LNA Array(v.18.0)(Exiqon).The results of mi RNA microarray analysis were further confirmed by real-time quantitative polymerase chain reaction(q PCR).The differentially expressed mi RNA target genes and signaling pathways involved were predicted by bioinformatics analysis.Results: Mi RNA microarray chip analysis revealed that a total of 297 specific mi RNA were selected from the necrotic group and the normal group,which had 35 up-regu lated genes and 262 down-regulated genes.Through the fold change and p-value screened expression is the difference of 2 up-regulated and 2 down mi RNAs,q PCR detection,hsa-mi R-93-5p,hsa-let-7i-5p in necrosis patients are more increased expression than normal,and on the contrary,hsa-mi R-122-5p,hsa-mi R-4711-3p in necrosis patients with expression decreased,which are consistented with microarray results.Using Target Scan,mir Base and mi Randa three online database to predict 2 upregulation and 2 down-regulation intersection target genes.Target genes of hsami R-93-5p was predicted of 103,and that of hsa-let-7i-5p was 107;and the target gene of hsa-mi R-122-5p was predicted of 31,and that of hsa-mi R-4711-3p was only prediction of 257 in the target Sran database,and the other two database prediction value is 0.Go analysis showed that cell metabolism,nervous system development,protein modification,intracellular protein modifications are closely related with the up-regulation mi RNAs.The down-regulation mi RNAs are closely related of the adjustable micro RNA and positioning,system development,neural development,organ development.Pathway analysis shows: the up-regulation mi RNAs combined with the protein to activate enzymes activity,the protein kinase,and down-regulation mi RNAs are related to protein and nucleic acid binding activity and ion channel.Conclusion: The differential mi RNA expression between TONFH and normal femoral are existence,which may be involved in the regulation of TONFH.The data of RT-PCR are consistented with the results of microarray scanning,and the further study on expression of mi RNA and its target m RNA will lay a foundation for further understanding of the molecular mechanisms involved in the pathogenesis of TIONFH in the process of repairing.