Correlation Between The Gene Single-nucleotide Polymorphism And Serum Concentration Of Tumor Necrosis Factor-alpha And Degenerative Lumbar Scoliosis

Objective: This study aim to investigate the correlation between tumornecrosis factor-alpha(TNF-α)gene single-nucleotide polymorphism and itsserum concentrations levels in degenerative lumbar scoliosis, which exploreetiology of degenerative lumbar scoliosis from genetic and molecular leveland provide the basis for analyzing on the etiology and prevention ofdegenerative lumbar scoliosis clinically. The definition of degenerative lumbarscoliosis is the occurrence of spinal deformity is based on lumbar degenerationafter skeletal maturity, with the coronal Cobb angle greater than10degrees,but not more than40degrees, which commonly occurred in more than50years people. Degenerative lumbar scoliosis is a three-dimensional andcomplex rotational deformity. With the progress of the study, such asbiomechanics and spinal anatomy, it is suggested that degenerative lumbarscoliosis not only has the vertebral imbalance on the coronal, but also has theloss of vertebral imbalance and vertebral rotation on the sagittal. A variety ofmalformations lead to low back pain and neurological symptoms of lowerextremity. At present, as our nation entering the aging society gradually, theincidence of this disease is increasing year by year. It has been an importantdisease that effects older people's quality of life. The etiology of degenerativelumbar scoliosis is very complex, and the pathogenesis is not clear. It isreported that there was correlation between TNF-α (Tumor necrosisfactor-alpha TNF-α) and degenerative lumbar scoliosis. TNF-α can mediateinflammatory response through the activation of inflammatory mediators, andcause lumbar degeneration. There is few relevant report about whether TNF-αgene polymorphism and its serum level is correlation with degenerativelumbar scoliosis. This study explored the correlation between gene polymorphism of the transcription initiation site upstream of TNF-α genepromoter308points and serum TNF-α level and degenerative lumbar scoliosisof the Han population of Northern Chinese, which detected by polymerasechain reaction-restriction fragment polymorphisms assay (PCR-RFLP) andenzyme linked immunosorbent assay (ELISA).Methods:60patients with degenerative lumbar scoliosis were selected aspatient group, who were from spinal surgery out-patient and wards in theThird Hospital of Hebei Medical University from October2009to December2011, the DLS group were all undertaken lumbar spine X-ray inanteroposterior and lateral and lumbar MRI. and select the same period in thehospital for60cases of healthy persons as normal control. All the cases wereall unrelated northern Chinese Han Pedigrees individuals, The two groupsmatched with gender, age, body mass index. Fresh peripheral blood of the twogroups were collected. DNA was extracted by the small amount of wholeblood Genomic DNA extraction reagent kit, specific fragments in TNF-α genepromoter region were amplified by polymerase chain reaction (PCR)technology, and then the amplified specific fragments were digested usingNcoI by restriction fragment length polymorphism (RFLP) technology, finallythe products were detected with a2%agarose gel electrophoresis and cases'genotype and allele frequency distribution were analyzed. Serum TNF-α levelswere determined by Enzyme-linked immunosorbent assay technique (ELISA).Using Adobe Photoshop6.0software, in DLS group the relative signalintensity of apical vertebra its upper and lower vertebral disc's nucleuspulposus and cerebrospinal fluid in T2-weighted MRI images were measured.DLS patients' Cobb angles were measured on the anteroposterior lumbar spineX-ray films. All the data were analyzed by Statistic Package for SocialScience (SPSS)13.0. Gender, body mass index and serum TNF-α levels werecompared using the t test or analysis of variance. The count data, such as age,genotype and allele frequency were compared using the χ2or Fisher exact test.Correlation between serum TNF-α levels and vertebral disc' degeneration andcorrelation between serum TNF-α levels and Cobb angles were made by Linearcorrelation analysis.Results: The comparison of the two groups' age,gender distribution andbody mass index did not show significant statistically, which indicated thatage, gender distribution, body mass index were matched. The amplificationTNF-α-308fragments was107bp by polymerase chain reaction and there werethree genotypes after digested by incision enzyme, including G/G genotype,G/A genotype, A/A genotype. On the electropherogram G/G genotype had twobright bars(87bp,20bp),G/A genotype had three bright bars(107bp,87bp,20bp), and A/A genotype had one bright bars(107bp). Three genotypesfrequencies of the two groups all accorded with Hardy-Weinberg equilibrium,so cases of the two groups were representative, In DLS group, the frequenciesof G/G genotype, G/A genotype and A/A genotype were70.0%,26.7%and3.3%, in the control group the frequencies of the three genotypes were86.7%,11.7%and1.7%. The three genotypes frequencies of the two groups had nosignificant differences. In DLS group the frequencies of G/G genotype andnon-G/G (G/A,A/A) genotype and the frequencies of allele G and A were70.0%,30.0%and83.3%,16.7%, in the control group were86.7%,13.3%and92.5%,7.5%. The comparion of two groups had significant difference.Compared with G/G genotype, non-G/G genotype had a higher risk fordeveloping degenerative lumbar scoliosis (OR=3.245,95%CI=1.240～8.495),Csaes with allele A, compared with allele G, had a higher risk for developingdegenerative lumbar scoliosis(OR=2.467,95%CI=1.074～5.667). CytokineTNF-α serum concentration in DLS group was163.25±3.44ng/L and incontrol group was51.12±0.92ng/L, The concentration of DLS group wassignificantly higher than that of control group, and there was significantdifference, and the patient group was significantly higher. Serum TNF-αconcentration has a negative correlation with the RSI and a positivecorrelation with the Cobb angle. According to the disc degeneration grade,DLS group was divided, there were significant differences between differentgroups.Conclusion: In Han population of Northern Chinese, TNF-α-308 polymorphism was no significant correlation with degenerative lumbarscoliosis. Serum TNF-α concentration in degenerative lumbar scoliosispatients was significantly increased. Serum TNF-α concentration has anegative correlation with the RSI and a positive correlation with the Cobbangle. The higher serum TNF-α concentration is, the more severe discdegeneration occur. TNF-α may play an important role in the degeneration ofdegenerative lumbar scoliosis.