云南省突发性耳聋SOD2基因遗传相关性研究

发布时间：2018-11-21 12:18

【摘要】：研究表明,突发性耳聋存在易感性差异,这种易感性差异可能与突发性耳聋的发病机制中有着重要生物学功能的基因有关。超氧化物歧化酶2(SOD2)是线粒体内唯一的抗氧化酶,可保护、对抗内耳免受自由基的损伤,其基因发生突变有可能影响超氧化物歧化酶2对内耳的保护作用,从而增加个体突发性耳聋的患病风险。本项目着眼于超氧化物歧化酶2基因的突变与突发性耳聋病因学的关系,主要运用基因组DNA的提取、多重SnaPshot等技术研究超氧化物歧化酶2(SOD2)基因的多个单核苷酸多态性(SNP)位点与突发性耳聋的关系,并可基于此制备基因组探针,从而发现具有潜在突聋危险性的病患,在临床上为分子诊断和个性化治疗提供理论依据。确定突发性耳聋的诊断标准诊断依据：1突然发生的,可在数分钟、数小时或三天以内。2非波动性感音神经性听力损失,可为轻、中或重度,甚至全聋。至少在相连的两个频率听力下降20db以上。多为单侧,或偶有双侧同时或先后发生。3病因不明(未发现明确原因或者包括全身或局部因素)。4伴耳鸣、耳堵塞感。5伴眩晕、恶心、呕吐,但不反复发作。6除了第八颅神经外,无其他颅神经受损症状。突发性耳聋血样采集程序和文件建立：由云南省第一人民医院入院病人提供的诊断标准,保证病例收集符合国内外流行病学调查规范并按同样标准执行。病史采集内容：采用云南省第一人民医院采血单,内容含入院病人的基本信息、耳科病史、糖尿病等其他疾病。由医生填写并保存。本课题共收集样本165例,其中80例突发性耳聋患者,85例正常控制组。并采集外周静脉血,通过采集肘静脉血2ml, EDTA抗凝保存于-80度冰箱保存,提取全基因组DNA、并通过SnaPshot方法对SOD2基因位点进行SNP分型。研究结果：在本次研究中,选取中国云南地区人群,一共165例中病人80,控制组85例,选取5个SNP位点(分别是rs5746136, rs5746129, rs2842960, rs4987023,rs4880),通过SnapShot基因分型,并用SPSS20.0进行卡方检验,发现突发性耳聋与SOD2基因的rs5746136位点存在易感性关系,其中AG基因型会赋予了突发性耳聋的风险(P=0.016; OR=2.136,95%CI=1.147-3.978)。其他位点并没有发现和突发性耳聋具有相关性。考虑到单倍型可能影响疾病的表型,于是我们在两个组间进行单倍型频率分析。由5个SNP位点进行构建单倍型,发现构建的单倍型在两组间无统计学差异。
[Abstract]:The study shows that there is a difference in susceptibility to sudden deafness, which may be related to genes with important biological function in the pathogenesis of sudden deafness. Superoxide dismutase 2 (SOD2) is the only antioxidant enzyme in mitochondria, which can protect the inner ear from the damage of free radicals. The mutation of its gene may affect the protective effect of superoxide dismutase 2 on the inner ear. This increases the risk of sudden deafness in individuals. This project focuses on the relationship between the mutation of superoxide dismutase 2 gene and the etiology of sudden deafness, and mainly uses the extraction of genomic DNA. Multiple SnaPshot techniques were used to study the relationship between multiple single nucleotide polymorphism (SNP) loci of superoxide dismutase 2 (SOD2) gene and sudden deafness. It provides theoretical basis for molecular diagnosis and individualized therapy in clinic. The diagnostic criteria for sudden deafness were determined as follows: (1) sudden onset may occur within a few minutes, hours or three days. 2 Non-fluctuating sensorineural hearing loss may be mild, moderate or severe, or even total deafness. At least at two consecutive frequencies hearing loss 20db above. Most of them were unilateral, or occasionally occurred at the same time or successively on both sides. 3 unknown etiology (no definite cause or including systemic or local factors). 4 tinnitus, ear clogging feeling. 5 with vertigo, nausea, vomiting. But no recurrent attacks. 6 except the eighth cranial nerve, no other cranial nerve damage symptoms. Establishment of procedures and documents for collecting blood samples of sudden deafness: diagnostic criteria provided by patients admitted to the first people's Hospital of Yunnan Province ensure that the collection of cases conforms to the epidemiological investigation norms at home and abroad and is carried out in accordance with the same standards. Collection of medical history: the first people's Hospital of Yunnan Province was used to collect blood, which contained the basic information of the hospitalized patients, the history of otology, diabetes and other diseases. Fill in and save by the doctor. A total of 165 samples were collected, including 80 sudden deafness patients and 85 normal control groups. The peripheral venous blood was collected, and the EDTA anticoagulant was stored in -80 degree refrigerator. The whole genome DNA, was extracted and the SNP typing of SOD2 locus was carried out by SnaPshot method. Results: in this study, 80 out of 165 patients were selected from Yunnan region of China, 85 from control group, 5 SNP loci (rs5746136, rs5746129, rs2842960, rs4987023,rs4880) were selected, and SnapShot genotyping was used. By using SPSS20.0 chi-square test, we found that sudden deafness was susceptible to rs5746136 locus of SOD2 gene, and the AG genotype was associated with the risk of sudden deafness (P0. 016). OR=2.136,95%CI=1.147-3.978) No other loci were found to be associated with sudden deafness. Considering that haplotypes may affect disease phenotypes, we performed haplotype frequency analysis between the two groups. Haplotypes were constructed from 5 SNP loci, and there was no statistical difference between the two groups.
【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R764

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