To Explore The Value Of "Preventive Treatment Of Disease" In Early Diagnosis Of Steroid-induced Osteonecrosis Of Femoral Head By Differential Expression Of MicroRNAs In Urine Exosome

BackgroundOsteonecrosis of femoral head is one of the most common and intractable diseases that seriously endanger people' s health.It has a high disability rate,which has caused great mental and economic burden to individuals and society.The occurrence of osteonecrosis of femoral head is related to the gene change,but there is no definite familial heredity.At present,magnetic resonance imaging is considered to be the most effective noninvasive detection method for the diagnosis of osteonecrosis of femoral head.Unfortunately,most of the patients' femoral heads have progressed to the late stage at MRI examination and the prognosis is still poor so far.What' s more,MRI tests are expensive and can' t be used for patients with metal plants,which limit the clinical value of MRI in early diagnosis of osteonecrosis of femoral head.By tracing the literatures,we found that exosomes on behalf of liquid biopsies are widely used in the early diagnosis of multiple cancers and microRNAs have been considered to play a role in human orthopedic metabolie disease.The detection method is the first non-interventional and repeatable to obtain the samples to be detected,and the results are objective and reliable.ObjectiveThe aim of our study is to identify potential biomarkers related to steroid-induced osteonecrosis of femoral head by analyzing microRNAs in urine exosome(noninvasive biopsy),and to diagnose steroid-induced osteonecrosis of femoral head in a noninvasive way.Through bioinformatics analysis,we try to find signaling pathways related to steroid-induced osteonecrosis of the femoral head and provide a new approach for the treatment of steroid-induced osteonecrosis of femoral head.MethodsA total of 9 cases of steroid-induced osteonecrosis of femoral head and hip osteoarthritis were treated in Department of orthopedics,the First Affiliated Hospital of Guangzhou University of Chinese Medicine from January 2017 to June 2017,eollected midstream urine and extracted total RNA of urine exosomes,which using mieroRNA-seq technology,screening the specific expression of differential mieroRNAs in steroid-induced osteonecrosis of femoral head.To further enlarge the sample size,real-time fluorescence quantitative PCR was used to verify microRNAs expression in 10 healthy individuals,10 patients with hip osteoarthritis and 30 patients with steroid-induced osteonecrosis of femoral head.Using the bioinformatics(GO analysis,KEGG analysis and protein-protein interaction network)to predict the possible targets and closely related signaling pathways of mieroRNA.ResultsCompared with hip osteoarthritis,steroid-induced osteonecrosis of femoral head of urine exosomes microRNA sequencing data including 447 microRNAs,the differential expression of microRNAs(P<0.05)are 103.There are 28 distinct differentially expressed microRNAs(P<0.01),including 15 up-regulated microRNAs and 13 down-regulated microRNAs.3 up-regulated microRNAs(microRNA-200b-3p,microRNA-206,microRNA-100-5p)and 2 down-regulated microRNAs(microRNA-320b and microRNA-3180-3p)were selected to verify the results of microRNA sequencing by qRT-PCR.We found that steroid-induced osteonecrosis of femoral head compared with hip osteoarthritis the relative expression of microRNA-200b-3p,microRNA-206 and microRNA-100-5p was significantly up-regulated(P<0.01)and the relative expression of microRNA-3180-3p was significantly down-regulated(P<0.01).MieroRNA-200b-3p,microRNA-206,microRNA-100-5p,and microRNA-3180-3p areas under the ROC curves are 0.9383,0.9259,0.8765 and 0.6111 in turn.Bioinformatics analysis and prediction of the top ten hub genes of microRNA-206 are AKT1,CHEK1,EGFR,BRCA1,CDK4,BCL2,VEGFA,CCND1,ESR1 and BUB1B.They may be involved in the signaling pathways such as Cell cycle,PI3K-Akt signaling pathway,Estrogen signaling pathway and FoxO signaling pathway.ConclusionThere are significant differentially expressed microRNAs in patients with steroid-induced osteonecrosis of femoral head of urine exosomes(micr oRNA-200b-3p?microRNA-206?microRNA-100-5p and microRNA-3180-3p),which c an serve as steroid-induced osteonecrosis of femoral head new biomarkers and give the "preventive treatment of disease" theory in the new connot ation of femoral head necrosis.The differential expression of microRNA-206 is likely to be involved in the pathogenesis of disease through PI3K-A kt,Estrogen and FoxO signaling pathways,providing a theoretical basis f or the limited prevention and treatment of disease.