Relationship Between MIF,TGFB,IFN? Gene Polymorphism And Spinal Tuberculosis Susceptibility And Expression In Intervertebral Disc And Its Clinical Data

1 Background and Purpose Tuberculosis has been in existence for thousands of years and is still the most important health problem in the world.Mortality in sputum smear-positive tuberculosis patients is 70%,tuberculosis is infected by Mycobacterium tuberculosis and causes young people as the main infected population A chronic infectious disease that seriously affects human health.However,this situation has not been controlled over time.In general,about 5% to 15% of the world's population(estimated to be between 20 and 30 billion)is infected with Mycobacterium tuberculosis in life and eventually leads to tuberculosis.However,the probability of tuberculosis in developing countries is higher than the probability of HIV infection,and its mortality rate is also higher than the death of HIV-related infectious diseases,is the main cause of infectious disease death.The incidence of tuberculosis is affected by a variety of factors,in which the host gene plays an important role in it.In developing countries,spinal tuberculosis is the most common extrapulmonary tuberculosis,spinaltuberculosis damage in patients with vertebral and intervertebral disc and the formation of flow abscess,often complicated by spinal deformity,paralysis,severe and even lead to death.In the pathogenesis of spinal tuberculosis,intervertebral disc destruction is a characteristic change of spinal tuberculosis disease,but its mechanism is not clear.In recent years,the study found that a variety of cytokines in the pathogenesis of tuberculosis play a more and more important role in the study has shown that MIF,TGF-?1,IFN-? and other cytokines play an important role in tuberculosis immunization,but the above three Whether cytokines play a role in the intervertebral disc disruption of patients with spinal tuberculosis requires more in-depth research.MIF can kill Mycobacterium tuberculosis in vitro,in the body's tuberculosis immunity can inhibit macrophages random walk to the invasion of Mycobacterium tuberculosis site and release a variety of cytokines and increase the inflammatory response,promote matrix metalloproteinases,NO,and IFN-?release,and matrix metalloproteinases also promote and exacerbate the occurrence of disc disruption,so MIF also has anti-tuberculosis immunity and proinflammatory effect.TGF-?1 can inhibit the activity of IFN-? and TNF-?,and up-regulate the expression of MMP-2 and MMP-9,inhibit the function of T lymphocytes and reduce the resistance to Mycobacterium tuberculosis and aggravate the destruction of intervertebral disc.IFN-? can kill Mycobacterium tuberculosis by activating T lymphocytes and effector cells and exert anti-tuberculosis immune function.At present,the study of MIF,TGF-?1 and IFN-? in the pathogenesis of spinal tuberculosis is rare,and its gene polymorphism and host genetic susceptibility research is also rare,especially on the above-mentioned cytokines in patients with spinal fusion In the pathogenesis of rare,based on the above cytokines in the role of tuberculosisimmunization,so we selected the above three kinds of cytokines to study,the study will help deepen our genetic susceptibility to patients with spinal tuberculosis and intervertebral disc damage Mechanism of understanding.2 Methods2.1 Control group: September 2014 to September 2015,our hospital physical examination center blood diagnosis confirmed the normal Guangxi Han people 110 cases.Normal inclusion criteria include: 1.Inclusions can not have immunological diseases such as rheumatism,rheumatism,Sjogren's syndrome,systemic lupus erythematosus,ankylosing spondylitis and so on.2.Except for tumor patients.3.Except for HIV-positive patients.4.Except for tuberculosis patients.Spine fracture group: September 2014 to June 2016 in our hospital spine surgery for the diagnosis of spinal fractures,the need for intervertebral disc resection surgery,a total of 10 cases(patients are normal for the Han nationality).Normal patients also need to meet the following requirements: 1.Patients can not have immunological diseases: such as rheumatism,rheumatoid,Sjogren's syndrome,systemic lupus erythematosus,ankylosing spondylitis.2.Except for tumor patients.3.Except for HIV-positive patients.4.Histopathological examination and imaging diagnosis are consistent with the normal intervertebral disc tissue.spinal tuberculosis group:Selected from September 2014 to June 2016 in our Hospital of Spine Bone Surgery for the diagnosis of spinal tuberculosis and surgical indications for removal of lesions,decompression,bone graft and other operations,a total of 110 patients Are the Han Chinese people).Spinal tuberculosis patients admitted to the standard at the same time have the following: 1.With typical tuberculosis symptoms,afternoon fever,night sweats,fatigue,weight loss and other symptoms.2.Those who have symptoms of nerve compression.3.Diagnosis ofblood samples in patients with tuberculosis diagnosis.4.Postoperative histopathological examination was diagnosed as spinal tuberculosis.5.Imaging examination and tuberculin test in line with tuberculosis diagnosis.6.Exclude other immune diseases,cancer and HIV infection and so on.7.exclude tuberculosis patients.2.2 Study on the extraction of MIF,TGF-?1 and IFN-?1 by blood genomic DNA extraction technique and RNA extraction technique and protein extraction technology,common PCR,reverse transcription,gene sequencing and fluorescence quantitative PCR and Western blotting.The expression levels of MIF,TGF-?1 and IFN-? in normal intervertebral disc and tuberculous intervertebral disc and the m RNA transcription level of MIF,TGF-?1 and IFN-?were normal The expression and difference of intervertebral disc tissue and tuberculous intervertebral disc tissue.3 Results3.1 The results showed that the genotype and C allele of MIF rs755622polymorphism(CC + GC)genotype and C allele,TGF-?1rs4803455 locus were significantly higher than those of healthy control group(P < 0.05).The results showed that the levels of MIF,TGF-?1 and IFN-? in the intervertebral disc of patients with spinal tuberculosis were significantly higher than those in the spine fracture group(P <0.05).The m RNA expression of TGF-?1 and IFN-? in the intervertebral disc was significantly higher than that in the spine fracture group(P <0.05),while the expression of MIF m RNA in the tuberculosis group was significantly lower than that in the spinal fracture Group(P <0.05).3.2 The results show that the expression of TGF-?1 and IFN-? m RNA in the intervertebral disc of patients with spinal tuberculosis is negatively correlated.The expression of MIF m RNA in patients with spinal tuberculosiswas not related to the clinical data.The mRNA expression level of TGF-?1 and IFN-? in patients with spinal tuberculosis was not related to the segment,sex and so on.There was a positive correlation between smoking habits and TGF-?1m RNA expression(p = 0.026).(P = 0.026,P = 0.019).The m RNA expression level of TGF-?1 was significantly higher than that of the control group(P<0.05),and the expression of TGF-?1 m RNA was significantly higher in VAS score than 8-10 VAS score at 0-4 points of mild pain,IFN-? m RNA expression levels in the VAS score of 0-4 points when mild pain was significantly higher than the VAS score at 8-10 points for severe pain.(P = 0.009,P = 0.022).The m RNA expression level of TGF-?1 was significantly higher than that of the control group(P <0.05),and the expression level of TGF-?1 m RNA was significantly higher than that of the control group Patients with IFN-? m RNA expression levels in patients with a duration of less than 3 months were significantly higher than those with a duration of 3 to 12 months.Serum CRP and ESR levels could affect the expression of TGF-?1 and IFN-? m RNA(CRP:P = 0.004,P = 0.011,ESR: P = 0.007,P = 0.026)and CRP> 30 mg / L,TGF-?1m RNA expression level was high,CRP + 10 mg / L,the expression level of IFN-? m RNA was high,ESRs> 40 mm / h,TGF-?1 m RNA expression level,ESRs <20 mm / h,The expression level of IFN-? m RNA is high.The higher the expression of TGF-?1 m RNA,the lower the expression of IFN-? m RNA(P<0.05).4 Conclusion4.1 In this study,we found that MIF rs755622 and TGF-?1 rs4803455 in Guangxi Han population were related to susceptibility to spinal tuberculosis,suggesting that allele C may be one of the factors that increase the susceptibility of spinal tuberculosis.4.2 In this experiment,we found that the levels of MIF,TGF-?1 and IFN-?in the intervertebral disc tissue of patients with spinal tuberculosis were expressed.MIF,TGF-?1 and IFN-? spinal tuberculosis patients in the intervertebral disc protein content is higher than the control group,so we speculate that the inflammatory reaction in the spinal tuberculosis process is a variety of different cytokines combined results,MIF,TGF-?1 may participate The immune process of spinal tuberculosis and mediate the pathological process of disc disruption,IFN-? may be involved in the immune process of spinal tuberculosis and play an important protective effect.4.3 In this experiment,it was confirmed that the expression level of TGF-?1 m RNA in the intervertebral disc tissue of patients with spinal tuberculosis was related to the amount of smoking in patients with spinal tuberculosis,suggesting that smoking may be an adverse factor affecting the prognosis of spinal tuberculosis.In this study,it was confirmed that the expression level of IFN-? m RNA in vertebral disc tissue was negatively correlated with VAS score,and the m RNA expression level of TGF-?1 was positively correlated with VAS score.Suggesting that TGF-?1 may cause pain aggravation by modulating the degree of inflammatory response.In this study,it was confirmed that the expression level of TGF-?1 m RNA in the intervertebral disc tissue of patients with spinal tuberculosis was positively correlated with the length of disease.The expression of IFN-? m RNA was negatively correlated with the duration of the disease.Suggesting that the length of the disease is closely related to the prognosis of patients with spinal tuberculosis.In this study,it was confirmed that the expression of TGF-?1 m RNA in the intervertebral disc tissue of patients with spinal tuberculosis was positively correlated with the level of erythrocyte sedimentation rate and C-reactive protein.The m RNAexpression level of IFN-? was negatively correlated with ESR and C-reactive protein.Suggesting that it plays a different role in the inflammatory reaction of spinal tuberculosis,and the effect and expression of TGF-?1 and IFN-? in different inflammatory stages are also different.