退变性腰椎侧凸TNF-α基因多态性和蛋白表达水平的相关研究  

The correlation between the polymorphism and protein expression level of tumor necrosis factor-alpha and degenerative lumbar scoliosis

作　　者：安志辉[1] 丁文元[1] 张剑刚[1] 杨少坤[1] 霍烈[1] 

机构地区：[1]河北医科大学第三医院脊柱外科,石家庄050051

出　　处：《中国骨与关节杂志》2012年第4期381-387,共7页Chinese Journal of Bone and Joint

基　　金：河北省自然基金资助项目(C2010000574)

摘　　要：目的研究探讨TNF-α基因单核苷酸多态性和血清中TNF-α水平与退变性腰椎侧凸的相关性。方法选取2009年10月至2011年12月于本院脊柱外科门诊及病房收治的60例退变性腰椎侧凸患者作为研究对象,均行腰椎正侧位X线片及腰椎MRI检查,选取同时期于该院进行健康体检者60例作为正常对照组,两组研究对象均系无血缘关系的北方汉族个体,在性别、年龄、体重指数与病例组相匹配。应用聚合酶链反应-限制性内切酶长度多态性(PCR-RFLP)技术对两组研究对象DNA标本TNF-α基因启动子区特异性片段进行扩增及对扩增的特异性片段用NcoI进行酶切,用酶联免疫吸附测定技术(ELISA),对两组研究对象血清TNF-α蛋白水平进行检测。利用Adobe Photoshop 6.0软件,测量病例组MRI图像中顶椎间盘及其上下椎间盘内髓核与脑脊液T_2加权像的相对信号强度及正位X线片上的Cobb角。结果两组研究对象各基因型频率分布符合Hardy-Weinberg遗传平衡定律,研究对象具有代表性。两组基因型频率差异无统计学意义(P>0.05)。G/G基因型频率与非G/G基因型频率(G/A、A/A)及等位基因G、A频率两组比较差异均有统计学意义(P<0.05)。相对危险度比较提示,非G/G基因型比G/G基因型患病的危险性增大,A等位基因携带者比非A等位基因携带者患病的危险性增加。细胞因子TNF-α血清浓度两组之间差异有统计学意义(P<0.05),且病例组显著高于对照组。TNF-α血清浓度与髓核相对信号强度呈负相关,与Cobb's角呈正相关。将病例组根据椎间盘退变等级分组,各组之间差异有统计学意义(P<0.05)。结论退变性腰椎侧凸患者血清中TNF-α浓度比正常人群血清水平明显增高,血清中TNF-α浓度与侧凸角呈正相关,与退变性腰椎侧凸椎间盘髓核信号强度呈负相关,血清TNF-α浓度越高,椎间盘退变程度越重,TNF-α是退变性腰椎侧凸发生发展的危险因素。Objective To investigate the correlation between tumor necrosis factor-alpha （TNF—α） gene single-nucleotide polymorphism and its serum concentration levels and degenerative lumbar scoliosis （DLS）. Methods 60 patients with DLS were selected as the patient group, who were from the Department of Spinal Surgery and wards in the Third Hospital of Hebei Medical University from October 2009 to December 2011. The ones of the patient group were all examined by anteroposterior and lateral X-rays of the lumbar spine and the lumbar MRI. 60 healthy cases were selected in the same period in the hospital as the normal control group. All the patients were unrelated northern Chinese Han Pedigrees. The 2 groups matched with respect to gender, age and body mass index. The specific fragments of the TNF—α gene promoter region of the 2 groups＇ DNA samples were amplified by polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） technique, and then the amplified specific fragments were digested using NcoI. The protein levels of serum TNF—α of the 2 groups were detected by enzyme-linked immunosorbent assay technique （ELISA） technique. Using Adobe Photoshop 6.0 software, in the patient group, the relative signal intensity of the apical vertebra and its upper and lower vertebral disc＇s nucleus pulposus and cerebrospinal fluid in T2-weighted MRI images were measured. The Cobb angle of patients with DLS were measured on the anteroposterior X-ray films.Results The distribution of genotype frequencies of the 2 groups all accorded with the Hardy-Weinberg equilibrium, so cases of the 2 groups were representative. The differences of the genotype frequencies between the 2 groups had no significant difference （P〉0.05）. The differences between G/G genotype frequencies and non-G/G genotype frequencies （G/A, A/A） and the allele frequencies G and A were statistically significant （P〈0.05）. Compared with patients with G/G genotype, patients with non-G/G genotype had a higher risk of deve

关 键 词：TNF-Α 核苷酸 多态性 退变性侧凸 腰椎 相关性 

分 类 号：R682.3[医药卫生—骨科学]