Correlation between lumbar disc degeneration and COL9A2 gene single nucleotide polymorphisms

doi:10.3969/j.issn.2095-4344.2013.09.026

Abstract

Abstract:

BACKGROUND: Studies have shown that lumbar disc degeneration is closely related to genes.
OBJECTIVE: To explore the relationship between COL9A2 gene single nucleotide polymorphisms (rsl2722877, rs3737820 and rs209914) and lumbar disc degeneration.
METHODS: 280 patients with lumbar degenerative disc disease were selected, and 268 age- and sex-matched patients without lumbar degenerative disc disease were selected as control group. All the patients came from Guangxi Zhuang Autonomous Region. The blood samples were collected from the patients with lumbar degenerative disc disease and without lumbar degenerative disc disease to extract the genomic DNA and to design PCR primers and TaqMan probe for the rsl2722877 rs3737820 and rs209914 sites of COL9A2 gene. The allele frequency and genotype frequency were analyzed with SPSS17.0 software.
RESULTS AND CONCLUSION: The differences of genotype and the allele frequency distribution of rsl2722877 rs3737820 and rs209914 sites were statistically significant (P < 0.05). The COL9A2 gene single nucleotide polymorphisms rsl2722877, rs3737820 and rs209914 were closely related with susceptibility of lumbar degenerative disc disease in Guangxi Zhuang population.

Key words: bone and joint implants, academic discussion of bone and joint, lumbar, intervertebral disc, degenerative diseases, COL9A2 gene, the Guangxi Zhuang population, lumbar disc degeneration, single nucleotide polymorphism, genetic polymorphism, translational medicine, disc disease

CLC Number:

R318

Chen Tao, Li Guan-bao, Liang Ke-you, Jia Shi-qing. Correlation between lumbar disc degeneration and COL9A2 gene single nucleotide polymorphisms[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(9): 1695-1702.

Figures/Tables 9

1 病例分析分析病例来自2002年3月1至10日广西玉林骨科医院就诊的280例腰椎间盘退变性疾病患者，男性136例，女性144例，年龄(53.8±6.9)岁；腰椎非椎间盘退变性疾病对照组，268例，其中男性127例，女性141例，年龄(52.8±4.2)岁；记录腰椎间盘退变性疾病组和腰椎非椎间盘退变性疾病对照组的所有资料。根据椎间盘退变Pfirrmann分级系统，观察病例组椎间盘退变的磁共振成像表现。髓核信号强度无或轻微下降、椎间盘高度正常为正常组；髓核信号强度与椎间盘高度均降低为病例组。病例组：纳入标准：①病史、体格检查、影像学符合腰椎间盘退变的诊断标准。②有完整的临床资料和影像学资料(包括X射线片、腰椎CT、腰椎MRI)。③广西壮族人群的散发性病例，中、重劳动强度。④知情同意。排除标准：①既往有脊柱外伤史、代谢性骨病史、下肢不等长、脊柱感染、肿瘤病史，及青少年时期诊断脊柱侧凸等脊柱畸形。②影像学资料、临床资料不完整的腰椎间盘退变患者。③未获得知情同意。正常组：纳入标准：①同期广西玉林骨科医院骨科门诊和体检中心的非腰椎间盘退变者。②年龄、性别、劳动强度与病例组匹配，均为广西壮族人群。③知情同意。排除标准：①未获得知情同意。②椎间盘源性腰痛病史、既往有相关脊柱外伤、手术史；脊柱肿瘤、结核等排除。 DNA提取：采用QIAampDNA Blood Mini kit (Qiagen公司)试剂盒法提取样本外周静脉血的全基因组DNA。2.5%琼脂糖凝胶电泳检测基因组DNA的完整性和纯度，-20 ℃储存待用。引物和探针设计：COL9A2基因序列来自Gen-bank。运用Primer Premier 6.0软件自行设计引物并应用oligo6.0软件评价，引物序列见表3-5。"

2 腰椎间盘退变与相关基因单核苷酸多态性的关系 2.1 资料来源检索数据库有关腰椎间盘退变与相关COL9A2基因单核苷酸多态性研究的文献[26-27]，检索时间范围在2006至2012年，检索词为“椎间盘；退行性病变；腰椎间盘退变；单核苷酸多态性；基因多态性”，检索到相关文献52篇，结果分析相关文献15篇[30-44]。 2.2 纳入标准 ①具有原创性，论点论据可靠的有关腰椎间盘退变与相关COL9A2基因单核苷酸多态性研究相关的基础和临床研究文献。②观点明确，分析腰椎间盘退变的表现、机制和影响因素研究相关文献。③文章主题内容与腰椎间盘退变发病机制研究联系紧密的相关文献。 2.3 排除标准 ①排除重复研究、普通综述或Meta分析类文章。②排除与文章目的无关的文献。③排除较陈旧的理论观点。 2.4 分析指标 ①COL9A2基因单核苷酸多态性与腰椎间盘退变相关性。②COL9A2基因单核苷酸多态性与腰椎间盘退变相关病例分析。③腰椎间盘退变相关基因单核苷酸多态性分析。 2.5 腰椎间盘退变与COL9A2基因单核苷酸多态性的文献分析腰椎间盘退变性疾病的候选基因均为与腰椎间盘结构、成分相关及成分代谢相关的基因。Ⅸ型胶原是由3条a链构成的异三聚体，分别由COL9A1、COL9A2和COL9A3基因编码。Ⅸ型胶原在人椎间盘内的细胞来源及合成与关节软骨相似，提示人椎间盘内Ⅸ型胶原具有类似功能。COL9A2 基因位于1号染色体，是编码Ⅸ型胶原蛋白a2链的基因。在退变的椎间盘中，Ⅸ型胶原基因表达停止或减少，导致蛋白多糖与Ⅸ型胶原之间的稳定关系不能维持，髓核内胶原纤维丝间黏附能力下降而变得松散，其纤维丝直径也发生了变化，从而髓核的生物力学完整性遭受破坏，促进腰椎间盘退变性疾病的形成[28]。动物实验提示表达有大片段阅读框内缺失的截短Ⅸ型胶原a链的转基因小鼠家系，证实这些纯合子和杂合子小鼠在6个月龄都出现关节软骨退变的改变，进一步的影像学和组织学分析显示小鼠发生了脊柱的病变，包括椎间盘膨出，轻微的骨赘形成和椎间盘退变加速[29]。COL9A2基因单核苷酸多态性与腰椎间盘退变的相关性的文献分析，见表9。"

References

[1] Deyo RA,Weinstein JN. Low back pain. N Engl J Med.2001; 344(5):363-370.

[2] 吕浩然,刘尚礼,丁悦,等.动物模型椎间盘退变全程基因变化的对比[J].中国矫形外科杂志,2005,13(11):846-849.

[3] Olmarker K,Rydevik B.Selective inhibition of tumor necrosis factor-alpha prevents nucleus pulposus-induced thrombus formation, intraneural edema, and reduction of nerve conduction velocity: possible implications for future pharmacologic treatment strategies of sciatica.Spine.2001; 26(8):863-869.

[4] Chan D,Song Y,Sham P,et al.Genetics of disc degeneration. Eur Spine J.2006;15(3):317-325.

[5] 程勇军,沈洪兴.椎间盘退变相关基因单核苷酸多态性的研究进展[J].第二军医大学学报,2011,32(11):1244-1248.

[6] 何志敏,陈德玉,郭永飞,等.腰椎间盘突出症再手术的相关因素及再手术方式分析[J].中国矫形外科杂志,2006,14(15):1130-1133.

[7] 百度百科.腰椎间盘[DB/OL].2012-10-11.http://baike.baidu.com /view/1740926.htm

[8] 朱若夫,孙树新.腰椎间盘突出症微创手术治疗的研究进展[J].中国矫形外科杂志,2004,12(17):1334-1336.

[9] Thompson JP,Pearce RH,Schechter MT,et al.Preliminary evaluation of a scheme for grading the gross morphology of the human intervertebral disc.Spine.1990;15(5):411-415.

[10] Bernard TN Jr.Lumbar discography followed by computed tomography. Refining the diagnosis of low-back pain. Spine. 1990;15(7):690-707.

[11] Ibrahim MA,Jesmanowicz A,Hyde JS,et al.Contrast enhancement of normal intervertebral disks: time and dose dependence.AJNR Am J Neuroradiol.1994;15(3):419-423.

[12] Videman T,Nummi P,Battié MC,et al.Digital assessment of MRI for lumbar disc desiccation. A comparison of digital versus subjective assessments and digital intensity profiles versus discogram and macroanatomic findings.Spine. 1994; 19(2):192-198.

[13] Antoniou J,Steffen T,Nelson F,et al.The human lumbar intervertebral disc: evidence for changes in the biosynthesis and denaturation of the extracellular matrix with growth, maturation, ageing, and degeneration.J Clin Invest.1996; 98(4):996-1003.

[14] 孔杰,胡有谷.腰椎间盘退变的遗传因素研究进展[J].中国脊柱脊髓杂志,2008,18(7):546-549.

[15] Smith L.Chemonucleolysis. Personal history, trials, and tribulations.Clin Orthop Relat Res.1993;Feb(287):117-124.

[16] Kohyama K,Saura R,Doita M,et al.Intervertebral disc cell apoptosis by nitric oxide: biological understanding of intervertebral disc degeneration.Kobe J Med Sci.2000; 46(6):283-295.

[17] Olmarker K,Rydevik B.Selective inhibition of tumor necrosis factor-alpha prevents nucleus pulposus-induced thrombus formation, intraneural edema, and reduction of nerve conduction velocity: possible implications for future pharmacologic treatment strategies of sciatica.Spine. 2001;26(8):863-869.

[18] Doita M,Kanatani T,Ozaki T,et al.Influence of macrophage infiltration of herniated disc tissue on the production of matrix metalloproteinases leading to disc resorption.Spine.2001; 26(14):1522-1527.

[19] Paassilta P,Lohiniva J,Göring HH, et al.Identification of a novel common genetic risk factor for lumbar disk disease. JAMA.2001;285(14):1843-1849.

[20] 姜莉.细胞因子在椎间盘退变中的作用[J].中国康复医学杂志, 2003,18(1):58-60.

[21] 赵勇,王文波.Sox9基因与椎间盘退变[J].中华外科杂志,2005, 43(8):544-545.

[22] 叶伟,马若凡,苏培强,等.IL-1β基因单核苷酸多态性与腰椎间盘疾病的相关性[J].遗传,2007,29(8):923-928.

[23] 唐颖,袁寒艳,王子平,等.基质金属蛋白酶-3和维生素D受体的基因多态性与腰椎间盘退变的易感性[J].复旦学报:医学版,2007, 34(1):37-41.

[24] 肖斌,田伟,赵丹慧,等.TIMP-1 666C＞T单核苷酸多态性与腰椎间盘退变相关性分析[J].中华医学杂志,2010,90(41):2939- 2942.

[25] 生物谷.单核苷酸多态性简介[DB/OL].2006-12-26. http://www.bioon.com/experiment/mob/256336.shtml

[26] 中国知网.中国学术期刊总库[DB/OL].2012-08-10. https://www.cnki.net

[27] SCI数据库.Web of Sciencevia ISI Web of Knowledge[DB/OL]. 2012-08-10.http://ip-science.thomsonreuters.com/mjl

[28] Solovieva S,Lohiniva J,Leino-Arjas P,et al.COL9A3 gene polymorphism and obesity in intervertebral disc degeneration of the lumbar spine: evidence of gene-environment interaction. Spine.2002;27(23):2691-2696.

[29] 李文选,武汉,姚松梅,等.椎间盘退变易感基因研究进展[J].国际骨科学杂志,2011,32(6):369-371.

[30] Annunen S,Paassilta P,Lohiniva J,et al.An allele of COL9A2 associated with intervertebral disc disease.Science. 1999; 285(5426):409-412.

[31] Jim JJ,Noponen-Hietala N,Cheung KM,et al.The TRP2 allele of COL9A2 is an age-dependent risk factor for the development and severity of intervertebral disc degeneration.Spine.2005;30(24):2735-2742.

[32] Seki S,Kawaguchi Y,Mori M,et al.Association study of COL9A2 with lumbar disc disease in the Japanese population.J Hum Genet.2006;51(12):1063-1067.

[33] Higashino K,Matsui Y,Yagi S,et al.The alpha2 type IX collagen tryptophan polymorphism is associated with the severity of disc degeneration in younger patients with herniated nucleus pulposus of the lumbar spine.Int Orthop.2007;31(1):107-111.

[34] Knoeringer M,Reinke A,Trappe AE,et al.Absence of the mutated Trp2 allele but a common polymorphism of the COL9A2 collagen gene is associated with early recurrence after lumbar discectomy in a German population.Eur Spine J.2008;17(3):463-467.

[35] 孙正明,刘淼,张银刚.腰椎间盘退变相关基因的研究[J].中华骨科杂志,2008,28(6):516-519.

[36] Ala-Kokko L.Genetic risk factors for lumbar disc disease.Ann Med.2002;34(1):42-47.

[37] 陈俊英.腰椎间盘退变与Ⅰ型胶原含量及基质金属蛋白酶-3的关系[D].辽宁省:中国医科大学,2007:1-46.

[38] Boos N,Nerlich AG,Wiest I,et al.Immunolocalization of type X collagen in human lumbar intervertebral discs during ageing and degeneration.Histochem Cell Biol.1997;108(6):471-480.

[39] 于斌,吴志宏,邱贵兴,等.中国北方人群维生素D受体基因多态性与腰椎退变性椎间盘疾病关联研究[J].中国骨与关节外科,2012, 5(2):154-158.

[40] Takahashi M,Haro H,Wakabayashi Y,et al.The association of degeneration of the intervertebral disc with 5a/6a polymorphism in the promoter of the human matrix metalloproteinase-3 gene.J Bone Joint Surg Br.2001;83(4): 491-495.

[41] 吴小涛,赵延勋.白介素-1α在退变腰椎间盘中的分布与神经根性疼痛的相关性[J].中国临床康复,2002,6(22):3352-3353.

[42] Gruber HE,Norton HJ,Ingram JA,et al.The SOX9 transcription factor in the human disc: decreased immunolocalization with age and disc degeneration.Spine.2005;30(6):625-630.

[43] Wrocklage C,Wassmann H,Paulus W.COL9A2 allelotypes in intervertebral disc disease.Biochem Biophys Res Commun. 2000;279(2):398-400.

[44] 宋海峰,吴志宏,闫家智,等.腰椎间盘退变性疾病与COL9A2基因多态性的关系[J].中华实验外科杂志,2011,28(8):1381-1383.