Transcript Levels Of Major MMPs And ADAMTS-4in Relation To The Clinicopathological Profile Of Patients With Tuberculous Intervertebral Discs And Healthy Controls

1Background and PurposeSpinal tuberculosis is the most common form of bone tuberculosis inChina. Poverty, malnutrition, HIV and drug resistance has accelerated thespread of tuberculosis of spine. Spinal tuberculosis may lead to seriouscomplications of spinal kyphosis and paraplegia. In the early stage ofspinal tuberculosis, tuberculous infection occurs frequently in the anteriorvertebral body. But with the development of disease, tuberculosisinfection often forward longitudinal ligament direction spread, eventuallyled to the adjacent multi vertebral tuberculosis have occurred. When theadjacent vertebral body was tuberculosis, intervertebral disc will alsodamage, degeneration and necrosis. Although intervertebral disk failureis an important disease characteristics of disease of spinal tuberculosis,but about the mechanism of tuberculosis destroyed intervertebral dischas been rarely reported. Recent studies have shown that matrixmetalloproteinases (matrix metal, loproteinases, MMPs) and I typeplatelet binding protein motif (TSP) depolymerizing protein likemetalloproteinase (a disintegrin like and metal loproteinase with thrombospondin type1motifs, ADAMTS) play an important role inextracellular matrix destruction. MMPs is a kind of protease familycontaining Zn2+, due to its decomposition components of extracellularmatrix, play an important role in the human body physiological tissueremodeling and immune regulation. ADAMTS is an Zn2+dependentsecretory metalloprotease, where ADAMTS-4is a matrix of cartilageaggrecan (aggrecan) degradation enzyme. ADAMTS-4has degradationfor a large number of aggrecan contained in articular cartilage, causingresistance to pressure and tension and other physical properties of thearticular cartilage was decreased. As the failure process of theintervertebral disc in patients with spinal tuberculosis, intervertebral disccartilage matrix is destroyed, according to the theory of ADAMTS-4isinvolved in the destruction of the disc intervertebral spinal tuberculosis.The analysis of ADAMTS-4, MMP-3,-8,-9,-13, the expression of-14mRNA transcription level in tuberculous intervertebral disc tissue,degenerative disc and normal disc tissues and differences, by Spearmanrank correlation analysis of the expression of MMPs pairing correlations,a study on the mechanism of intervertebral disc injury that patients withspinal tuberculosis, which provides a theoretical basis for the treatment ofspinal tuberculosis. The study also through the detailed survey of clinicaldata of all patients with spinal tuberculosis, the combination of MMPs andADAMTS-4gene expression data, analysis of the gene expressioncorrelation between clinical data index and spinal tuberculosis, provide atheoretical basis for the comprehensive treatment of spinal tuberculosisdisease. 2Methods2.1spinal tuberculosis group (TB group):60cases of tuberculosis ofintervertebral disc tissue specimens were obtained from the FirstAffiliated Hospital of Guangxi Medical University from March1,2011toApril30,2012to accept operation treatment in spinal bone surgerypatients with spinal tuberculosis. Protrusion of the intervertebral discgroup (ID group):60cases of degenerative intervertebral disc tissuespecimens were obtained from the First Affiliated Hospital of GuangxiMedical University from March1,2011to April30,2012to acceptoperation in the treatment of spine bone surgery patients with protrusionof intervertebral disc. The normal group (control group):60cases ofpatients with normal intervertebral disc tissue specimens were obtainedfrom the First Affiliated Hospital of Guangxi Medical University fromMarch1,2011to April30,2012to accept operation treatment in spinalbone surgery in traumatic spinal fracture. After the experimental samplewas obtained, the tissue specimens were examined by two experiencedpathologists, ensure the accuracy of the sample. All samples wereobtained by surgical operation, immediately placed in liquid nitrogen andstored frozen at-80℃until RNA extraction.2.2using RNA extraction technology and real-time fluorescencequantitative PCR (Real time fluorescent quantitative PCR, RT-PCR)technology, research and analysis of the ADAMTS-4, MMP-3,-8,-9,-13,the expression of-14mRNA transcription level in tuberculousintervertebral disc tissue, degenerative disc and normal disc tissues anddifferences. By Spearman rank correlation analysis of MMPs expressionin the pairing correlation.2.3using the clinical questionnaire and laboratory examination method of combining the related data collection. This study selected clinical andpathological data of three groups including: age, sex, occupation,smoking habits, intensity of pain (visual analogue scale, VAS), spinaldisease, disease duration, C-reactive protein (CRP) and erythrocytesedimentation rate (ESR). The combination of MMPs and ADAMTS-4gene expression data, gene expression analysis of the correlationbetween clinical index and spinal tuberculosis3Results3.1RT-PCR showed, spinal tuberculosis group and protrusion of theintervertebral disc of ADAMTS-4expressions were significantly increased,compared with the normal group, the difference was statisticallysignificant (P<0.05). The expression of ADAMTS-4in spinal tuberculosisgroup although intervertebral disc herniation group, but the differencewas not statistically significant. Tuberculosis of the spine andintervertebral disc herniation group increased expression of MMP-3,compared with the normal group, the difference was statisticallysignificant (P<0.05). The expression of MMP-3in spinal tuberculosisgroup although intervertebral disc herniation group, but no significantdifference. Tuberculosis of the spine of the MMP-8expression wassignificantly increased, compared with the normal group, the differencewas statistically significant (P<0.05). The expression of MMP-8in spinaltuberculosis group than the disc herniation group, the difference wasstatistically significant (P<0.05). Tuberculosis of the spine of the MMP-9expression was significantly increased, compared with the normal group,the difference was statistically significant (P<0.05). The expression ofMMP-9in spinal tuberculosis group than the disc herniation group, the difference was statistically significant (P<0.05). Spinal tuberculosis groupand protrusion of the intervertebral disc of MMP-13expressions weresignificantly increased, compared with the normal group, the differencewas statistically significant (P<0.05). The expression of MMP-13in spinaltuberculosis group than the disc herniation group, the difference wasstatistically significant (P<0.05). Spinal tuberculosis group and protrusionof the intervertebral disc of MMP-14expressions were significantlyincreased, compared with the normal group, the difference wasstatistically significant (P<0.05). The expression of MMP-14in spinaltuberculosis group than the disc herniation group, but the difference wasnot statistically significant.3.2data showed that, the higher the level of disease more serious spinaltuberculosis patients with MMPs expression. Patients with severe spinaltuberculosis in MMP-3,-8,-9,-14expression level was significantlyincreased (P<0.05). The expression level of patients with severe spinaltuberculosis in MMP-13light was significantly increased (P<0.05).3.3spinal tuberculosis group and protrusion of the intervertebral disc ofthe MMPs expression has multiple correlation. Specifically, spinaltuberculosis group, the expression level of MMP-13and MMP-8,-9hadsignificant positive correlation. In addition, spinal tuberculosis in groupMMP-8, there was significant positive correlation between the expressionlevel of-9.3.4spinal tuberculosis group and protrusion of the intervertebral discgroup: ADAMTS-4gene expression and patient age, gender, occupation,smoking habits, pain intensity, the prevalence of spinal segment, diseaseduration, C-reactive protein and blood submerged mean no correlation (P>0.05). Spinal tuberculosis group: MMP-3expression has positivecorrelation with smoking habit (P=0.007). MMP-3gene expression andpatient age, gender, occupation, pain intensity, the prevalence of spinalsegment, disease duration, C-reactive protein and blood submergedmean no correlation (P>0.05). Spinal tuberculosis group: MMP-8geneexpression and age (P=0.011), pain intensity (P=0.014), C-reactiveprotein (P=0.004) and erythrocyte sedimentation rate (P=0.002) haspositive correlation. MMP-8gene expression and patient’s sex,occupation, the prevalence of spinal segment, disease duration, nocorrelation (P>0.05). Intervertebral disc herniation group: MMP-8geneexpression and age (P=0.032), C-reactive protein (P=0.003) anderythrocyte sedimentation rate (P=0.005) has positive correlation. Theexpression of MMP-8gene in patients with gender, occupation, smokinghabits, pain intensity, the prevalence of spinal segment, disease duration,no correlation (P>0.05). Patients with severe pain (VAS8-10) MMP-8levels than patients with mild pain (VAS0-4) increased significantly(P<0.05). Patients with high CRP (>30mg/L) MMP-8levels than patientswith low CRP (<10mg/L) increased significantly (P<0.05). Patients withhigh ESR (>40mm/h) MMP-8levels than patients with low ESR (<20mm/h) increased significantly (P<0.05).Spinal tuberculosis group: theexpression of MMP-9gene and the pain intensity (P=0.026), diseaseduration (P=0.021), C-reactive protein (P=0.006) and erythrocytesedimentation rate (P=0.013) has positive correlation. No correlationbetween MMP-9gene expression and patient age, gender, occupation,the prevalence of spinal segments (P>0.05). Patients with severe pain(VAS8-10) MMP-9levels than patients with mild pain (VAS0-4) increased significantly (P<0.05). Expression levels of3–12monthsdisease duration in patients with MMP-9than <3months disease durationwere significantly increased (P<0.05). Patients with high CRP (>30mg/L)MMP-9levels than patients with low CRP (<10mg/L) increasedsignificantly (P<0.05). Spinal tuberculosis group: the expression ofMMP-13gene and smoking habits (P=0.019), pain intensity (P=0.016),disease duration (P=0.012), C-reactive protein (P=0.002) anderythrocyte sedimentation rate (P=0.009) has positive correlation. MMP-9gene expression and patient age, gender, occupation, the prevalence ofspinal segments were not correlated (P>0.05). See table3-6for thespecific numerical. Patients with severe pain (VAS8-10) MMP-13levelsthan patients with mild pain (VAS0-4) increased significantly (P<0.05).Expression levels of3–12months disease duration in patients withMMP-13than <3months disease duration were significantly increased(P<0.05). Patients with high ESR (>40mm/h) MMP-13levels thanpatients with low ESR (<20mm/h) increased significantly (P<0.05).Group and lumbar spinal tuberculosis disc group: MMP-14geneexpression and patient age, smoking habits, gender, occupation, painintensity, the prevalence of spinal segment, disease duration, C-reactiveprotein and blood submerged mean no correlation (P>0.05).4Conclusion4.1this study confirmed the expression and intervertebral disctuberculous destruction of the ADAMTS-4intervertebral disc tissue, playan important role in the pathogenesis of speculation ADAMTS-4intervertebral disc destruction of spinal tuberculosis. Studies ofdegenerative lumbar disc tissue ADAMTS-4expression was significantly increased, ADAMTS-4played an important role in the pathogenesis ofintervertebral disc herniation.4.2this study confirmed that MMP-3, intervertebral disc tissue-8,-9,-13,-14expression and disc tuberculous destruction, suggesting that MMP-3,-8,-9,-13, play an important role in the pathogenesis of-14in theintervertebral disc damage in spinal tuberculosis. Studies of degenerativelumbar disc tissues of MMP-3,-13,-14expression was significantlyincreased, MMP-3,-13,-14play a very important role in the pathogenesisof intervertebral disc herniation.4.3this study confirmed that MMP-8,-9,-13in the intervertebral disc oftuberculous destroyed in the process played a synergistic role. The studyconfirmed that MMP-3,-8,-9,-14expression has correlation withintervertebral disc tuberculous destruction severity.4.4this study confirmed that MMP-3tuberculous intervertebral disctissues, there was a positive correlation between the expression of-13and smoking habits, smoking is that affect the prognosis of spinaltuberculosis. This study showed that MMP-8, tuberculous intervertebraldisc tissue-9,-13expression has positive correlation with pain intensity,pain may be through the way that increased inflammation, aggravation ofspinal tuberculosis patients. This study showed that MMP-9of tubercularlumbar disc tissues, expression and symptom duration of-13hadcorrelation, speculated that the symptom duration is related factorsaffecting the prognosis of spinal tuberculosis. This study showed thatMMP-8, tuberculous intervertebral disc tissue in-9, there was a positivecorrelation between the expression of-13and CRP, that high levels ofCRP affect the prognosis of spinal tuberculosis. This study showed that MMP-8, tuberculous intervertebral disc tissue in-9, there was a positivecorrelation between the expression of-13and ESR, that high levels ofESR affect the prognosis of spinal tuberculosis.