Study On The Changes Of MicroRNA Expression Profile In Peripheral Blood Of Spinal Tuberculosis And Its Potential Clinical Application

ObjectiveChanges in the microRNA expression profile in peripheral blood of spinal tuberculosis were detected,and microRNA related to toll-like receptor-mediated signal transduction pathway with large differential expression multiples were screened out to preliminarily explore the regulation mechanism of microRNA on spinal tuberculosis.Screening of microRNA with certain potential value in the differential diagnosis between spinal tuberculosis and brucellosis spondylitis.Confirmed from the gene level that ultra-short course chemotherapy less than 6 months after complete lesion clearance was feasible.MethodsUse the PAXgene Blood RNA Tube collect 5 cases of normal healthy volunteers and 5cases in ningxia medical university general hospital spinal orthopaedic spinal tuberculosis patients hospitalized with peripheral venous Blood,extraction of microRNA,using microRNA chip testing technology analyzed,and the screen microRNA of difference ratio less than 0.25,use TargetscanHuman 7.1 predicted microRNA target genes,Furthermore,microRNA that may be related to toll-like receptor-mediated signal transduction pathways were selected based on their target genes.Real-time quantitative PCR(qRT-PCR)was applied to verify the differentially expressed microRNA in venous blood samples of the spinal tuberculosis group and brucellosisspondylitis group(5 cases each),and to screen the differentially expressed mirnas.Sixty cases in the normal control group,81 cases in the spinal tuberculosis group and 52 cases in the brucellosis spondylitis group were collected for clinical large sample verification,and their potential value in the differential diagnosis between spinal tuberculosis and brucellosis spondylitis was evaluated by receiver operating characteristic curve analysis.Detect healthy volunteers and patients with spinal tuberculosis hospital after surgery and lesions cleared completely different treatment stages of peripheral blood(That is,at admission,at the end of postoperative chemotherapy and 1 year follow-up)specimens in the change of specific microRNA,from the level of gene confirmed completely lesions removal surgery less than 6 months of dots is feasible.ResultsComparison of microRNA expression profiles in peripheral venous blood of 5 normal healthy volunteers and 5 patients with spinal tuberculosis showed a total of 103 differentially expressed microRNA,102 of which were down-regulated and 1 of which was up-regulated.There were 6 microRNA with differential expression multiples less than 0.25 that might be related to toll-like receptor-mediated signaling pathways:hsa-mir-150-5p,hsa-mir-197-3p,hsa-mir-146a-5p,hsa-mir-6511a-3p,hsa-mir-342-3p,and hsa-mir-4732-3p.Five patients from the spinal tuberculosis group and brucellosis spondylitis group were selected for clinical small sample verification,the results showed that the expression of hsa-mir-150-5p in the spinal tuberculosis group was significantly reduced,the difference was statistically significant,the other five microRNA express no statistically significant difference in two groups,further clinical large sample verification,the results shown:hsa-miR-150-5-P expressed in spinal tuberculosis group decreased,with brucellosis spondylitis group and normal control group;No statistical difference in the expression of hsa-mir-150-5p between brucellosis spondylitis group and the normal control group.Receiver operating characteristiccurve was used to analyze the potential differential diagnosis value of hsa-mir-150-5p in the spinal tuberculosis group and brucellosis spondylitis group,and the area under the curve was0.831,and the Jordan index was the maximum value of 0.501,the sensitivity was 0.538,and the specificity was 0.963.Real-time quantitative PCR was used to detect the relative expression of hsa-mir-150-5p in the control group and spinal tuberculosis patients at the time of admission,at the end of postoperative chemotherapy,and at 1 year of postoperative follow-up.After statistical analysis,the relative expression of hsa-mir-150-5p was down compared with the normal control group at the time of admission of spinal tuberculosis,the reduce was statistically significant(P values< 5%).No difference in expression of hsa-mir-150-5p between the control group and the group at the end of postoperative chemotherapy and 1 year postoperative follow-up(p > 0.05).ConclusionCompared with the normal control group,microRNA expression profiles in peripheral blood samples of spinal tuberculosis patients were changed.,proving that microRNA plays a regulatory role in the occurrence and development of spinal tuberculosis.Among them,hsa-mir-150-5p,hsa-mir-197-3p,hsa-mir-146a-5p,hsa-mir-6511a-3p,hsa-mir-342-3p and hsa-mir-4732-3p may regulate the occurrence and development of spinal tuberculosis through toll-like receptor-mediated signal transduction pathway.Small clinical samples and large samples confirmed that hsa-mir-150-5p in peripheral blood has certain potential differential diagnosis value for spinal tuberculosis and brucellosis spondylitis.Compared with the normal control group,the expression of hsa-mir-150-5p in the peripheral blood of spinal tuberculosis patients on admission was down-regulated.Compared with the normal control group,there was no statistical difference in the expression of hsa-mir-150-5p in the peripheral blood at the end of postoperative chemotherapy and the yearof follow-up,it provide the basis of gene level for the feasibility study of ultra-short range chemotherapy less than 6 months on the basis of radical lesion removal.