Proteomic Screening Of LBP And Its Mediated TLRs/NF-kB Pathway In The Pathogenesis Of Spinal Tuberculosis

Objective: To screen differentially expressed proteins by proteomics technology in peripheral plasma of patients with spinal tuberculosis and explore its role in the pathogenesis of STB and its value as a potential diagnostic marker in the diagnosis of spinal tuberculosis.Methods: The peripheral plasma of 3 patients with spinal tuberculosis and 3 healthy persons were selected as the experimental group and the control group,respectively.The differential proteins in the experimental group were screened and identified by using proteomic isotope labeling relative and absolute quantitative techniques and liquid chromatography-mass spectrometry analysis techniques;Expand the sample size,select the peripheral blood of 30 patients with spinal tuberculosis as the experimental group and 30 healthy people with normal physical examination indexes as the control group,and carry out real-time quantitative reverse transcription polymerase chain reaction(RT-PCR)and enzyme-linked immunosorbent assay(ELISA)to verify their differential genes and proteins;GO and KEGG were enriched with biological function analysis software to analyze the function of differential proteins and the related signal pathways in the pathogenesis of spinal tuberculosis.Collect the clinical medical records of 100 patients with spinal tuberculosis who were admitted to the Department of Spinal Orthopedics of the General Hospital of Ningxia Medical University from May 2017 to May 2020(including the patient’s name,sex,age,laboratory examination,including blood routine,biochemistry,C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),acid-fast staining mycobacterium tuberculosis test,T-cell spot test for tuberculosis infection(T-SPOT),bacteriological culture,imaging examination and pathological examination;The peripheral blood of 100 healthy people with normal peripheral blood and physical examination indexes were tested by ELISA to verify the differential protein LBP of patients with spinal tuberculosis;T test,Spearman analysis,linear regression and ROC curve analysis were used to analyze the value of LBP in the diagnosis of spinal tuberculosis in peripheral blood;Use human monocyte THP-φ PMA was added to induce human macrophages,and BCG was added to simulate Mycobacterium tuberculosis(MTB)to infect human macrophages.After adding LBP small interference,the effect and change of LBP on inflammatory factors or bodies in the TLR signal pathway of spinal tuberculosis were verified,and the relationship between LBP and the TLRs/NF-κB related signal pathway related to the pathogenesis of spinal tuberculosis was clarified.Human mononuclear cells THP-φ were added into the Pharmaceutical Manufacturers Association(PMA)to induce human considerate parietal cells.By adding BCG to simulate Mycobacterium tuberculosis(MTB),we can infect adherent cells and differentiate them into macrophages.By adding small interference RNA of LBP,we can verify the effect of LBP on inflammatory factors or bodies in the TLRs/NF-κB signaling pathway of spinal tuberculosis.Furthermore,the relationship between LBP and TLRs/NF-κB signaling pathways related to spinal tuberculosis was elucidated.Results: 1.A total of 16 differential proteins were screened from the peripheral plasma of patients with spinal tuberculosis,of which 11 proteins were up-regulated,including Lipopolysaccharide-binding protein,Protein S100-A8,Protein S100-A9,Serum amyloid A-2protein,Leucine-rich alpha-2-glycoprotein,Proteoglycan 4,Complement factor I,Alpha-1-antitrypsin,Cathepsin D,Ceruloplasmin,Vinculin and 5 proteins down-regulated,including Lambda-Immunoglobulin4-60,Immunoglobulin heavy variable 3-64 D,Immunoglobulin kappa variable 2D-29,Non-functional immunoglobulin6D-41,Immunoglobulin heavy variable 1-45.2.The gene expression of LBP in the peripheral plasma of patients with spinal tuberculosis was 1.63 ± 0.31,and the protein expression was 7.57 ± 2.52ug/mL,which was significantly higher than that of the control group.3.In the laboratory examination of peripheral blood of patients with spinal tuberculosis,LBP detection value was significantly correlated with erythrocyte sedimentation rate and C-reactive protein detection value(P<0.001).4.In the laboratory examination of peripheral blood of patients with spinal tuberculosis,LBP detection value was significantly positively correlated with erythrocyte sedimentation rate and C-reactive protein detection value.LBP scores were 0.677(P<0.01),0.707(P<0.01),0.751(P<0.01),0.714(P<0.01),0.656(P<0.05).5.LBP affects TLR4/NF-κB Expression of related proteins in pathway.Conclusions: 1.Proteomics technology can be used to screen STB peripheral blood differential proteins.LBP is significantly highly expressed in the peripheral blood of patients with spinal tuberculosis,and its function is mainly related to biological functions such as inflammation,autophagy and chemokines.2.The differential expression of LBP in the peripheral blood of patients with spinal tuberculosis is significantly related to the examination indicators for clinical diagnosis of spinal tuberculosis,which can become a potential diagnostic marker of spinal tuberculosis.3.LBP can pass TLR4/NF-κ B signal pathway in the pathogenesis of spinal tuberculosis activates inflammatory factors or inflammatory bodies to produce inflammatory effects.