β-防御素（DEFB4）基因拷贝数变异与汉族强直性脊柱炎关联性研究

发布时间：2018-06-02 09:47

本文选题：强直性脊柱炎 + DEFB4　；参考：《安徽医科大学》2015年硕士论文

【摘要】：目的1.探讨β-防御素(DEFB4)基因拷贝数变异(Copy Number Variation,CNV)与强直性脊柱炎(Ankylosing Spondylitis,AS)遗传易感性之间的关联性;2.探讨基因CNV是否会影响疾病的严重程度,同时研究与基因CNV在疾病发生发展进程中相互影响的环境因素,为进一步探索AS的发病原因,阐明AS的发病机制提供重要线索和依据。方法1.2011~2014年,收集安徽医科大学第一附属医院风湿免疫科门诊确诊的AS患者作为病例组,同时期按性别、年龄成组匹配,来安徽医科大学第一附属医院体检中心体检的健康者为对照组,采用面对面问卷调查法,用课题组自行设计的调查问卷,收集患者和对照者的人口学资料:年龄、性别、文化水平、个人的行为习惯(吸烟、饮酒等)和饮食偏好,以及患者的临床资料:病程、症状持续时间、BASDAI疾病活动指数和BASFI功能指数;实验室血清学指标:血沉、C反应蛋白和HLA-B27等。在调查的同时采集患者和对照者的5ml外周血。2.DNA提取采用修改后的盐析法,选取具有高度重复区域的两个DEFB4基因片段(DEFB4_1和DEFB4_2),拷贝数的检测采用Accu CopyTM法。3.采用病例对照以及单纯病例研究设计,分析比较DEFB4基因中CNV在AS患者和对照者中的差异,采用logistic回归模型分析基因CNV与疾病、疾病的严重程度间的关联性,用单纯病例研究方法分析CNV与环境的交互作用。4.数据录入软件为Epidata3.1,采用双人录入并进行一致性检验,数据分析采用SPSS19.0统计软件实现,P0.05差异有统计学意义。结果1.共纳入806例研究对象,病例组405人,男性338人,平均年龄为27.91±8.93岁,中位病程为2.08年,HLA-B27阳性率为64%;对照组401人,男性324人,平均年龄为26.89±7.12岁。两组研究对象在性别(χ2=1.13,P=0.288)和年龄(t=1.77,P=0.076)上差异无统计学意义。2.DEFB4基因拷贝数的分布:AS患者,基因拷贝数范围为1~8,DEFB4_1主要为2~4拷贝,DEFB4_2主要为2~5拷贝。对照组拷贝数范围和病例组相同,DEFB4_1和DEFB4_2最多的拷贝均为3拷贝,分别占34.34%和27.14%。DEFB4_1(Z=0.63,P=0.528)和DEFB4_2(Z=1.27,P=0.111)拷贝数分布在病例组和对照组中差异无统计学意义。3.以3拷贝为界分两组分析,DEFB4_2拷贝数在病例组和对照组中的分布差异有统计学意义(χ2=4.49,P=0.034),高拷贝数组(≥4)人群是低拷贝数组(≤3)人群可能患AS风险的0.74倍,95%CI(0.56,0.98)。进一步分为三组:低拷贝数组(≤2),中拷贝数组(3),高拷贝数组(≥4),病例组和对照组拷贝数分布差异有统计学意义,χ2=8.68,P=0.013;与3拷贝数组比较,低拷贝数组和高拷贝数组人群分别是中拷贝数组人群可能患AS风险的0.68和0.62倍,但调整年龄和性别后,低拷贝数组与中拷贝数组比OR=0.69,95%CI(0.47,1.03),P=0.067,降低患AS风险的可能性无统计学意义。4.DEFB4_1和DEFB4_2拷贝数的增加是疾病严重度(BASDAI)增加的保护因素,OR,95%CI分别为0.71(0.56,0.90)和0.75(0.60,0.94),调整年龄、性别、病程之后,影响改变不大。未发现DEFB4基因CNV与身体功能(BASFI)的关联性。5.DEFB4_1:食盐口味中等的病人与食盐口味轻的病人相比,基因×环境交互作用OR=0.51,95%CI(0.28,0.94),P=0.032,其他环境因素并未发现与基因的交互作用。DEFB4_2未发现拷贝数变异与环境的交互作用。结论DEFB4基因高拷贝数(≥4)是AS发病的保护因素,并且独立于年龄和性别;基因CNV的拷贝数增加是独立于年龄、性别和病程对疾病严重度增加的保护因素,但未发现基因CNV的增加与患者身体功能状况的关联性;基因环境交互作用分析提示,饮食口味可能与DEFB4基因拷贝数变异共同影响患AS的风险。
[Abstract]:Objective 1. to explore the correlation between the genetic susceptibility of Copy Number Variation (DEFB4) gene copy number variation (CNV) and ankylosing spondylitis (Ankylosing Spondylitis, AS), and 2. to explore whether the gene CNV will affect the severity of the disease and study the environmental factors that interact with the co gene CNV in the progression of the disease, In order to further explore the causes of AS, clarify the pathogenesis of AS, provide important clues and basis. Method 1.2011~2014, collected AS patients in the Department of Rheumatology of the First Affiliated Hospital of Medical University Of Anhui as case group, and matched by sex and age group at the same time, to come to the physical examination center of the First Affiliated Hospital of Medical University Of Anhui for physical examination. In the control group, we used a face-to-face questionnaire to collect the demographic data of patients and controls: age, sex, cultural level, personal behavior habits (smoking, drinking, etc.) and dietary preference, and the clinical data of patients: duration of disease, duration of symptoms, and BASDAI disease activity index. And BASFI function index; laboratory serological index: erythrocyte sedimentation, C reactive protein and HLA-B27. The 5ml peripheral blood.2.DNA extraction of patients and controls was collected by modified salting out method, and two DEFB4 gene fragments (DEFB4_1 and DEFB4_2) with high repetition area were selected, and the detection of copy number was carried out by Accu CopyTM method.3. mining Use case control and simple case study design, analyze and compare the difference of CNV in AS patients and controls in DEFB4 gene, use logistic regression model to analyze the correlation between gene CNV and disease, the severity of disease, and analyze the interaction of CNV and environment by simple case study method,.4. data entry software is Epidata3.1. The data analysis was realized with SPSS19.0 statistical software, and the difference of P0.05 was statistically significant. Results 1. a total of 806 subjects were included in the study. The case group was 405, male 338, the average age was 27.91 + 8.93, the median course was 2.08 years and the HLA-B27 positive rate was 64%; the control group was 401, 324 men and average age. There was no statistically significant difference in the distribution of.2.DEFB4 genes in sex (x 2=1.13, P=0.288) and age (t=1.77, P=0.076) in two groups of subjects: AS patients, 1~8, DEFB4_1 mainly 2~4 copies, DEFB4_2 mainly 2~5 copy, and the same number of copies of the control group as the case group, DEFB4_1, and cases. The most copies of the copies were 3 copies, which accounted for 34.34% and 27.14%.DEFB4_1 (Z=0.63, P=0.528) and DEFB4_2 (Z=1.27, P=0.111) copy number distribution in the case group and the control group with no statistically significant difference.3. with 3 copies as the two groups. The distribution difference of DEFB4_2 in the case group and the contrast group was statistically significant (x 2=4.49, P=0.034). High copy array (> 4) population is low copy array (less than 3) people may suffer 0.74 times the risk of AS, 95%CI (0.56,0.98). Further divided into three groups: low copy array (< < 2), medium copy array (3), high copy array (> 4). The difference of copy number distribution in case group and control group is statistically significant, X 2=8.68, P=0.013; compared with 3 copy array, low copy Array and high copy array groups are 0.68 and 0.62 times the risk of AS, respectively, but after adjusting age and sex, low copy array and medium copy array are more than OR=0.69,95%CI (0.47,1.03), P=0.067, and the possibility of reducing AS risk is not statistically significant.4.DEFB4_1 and DEFB4_2 copy number increase is disease severity (BAS). DAI) increased protective factors, OR, 95%CI were 0.71 (0.56,0.90) and 0.75 (0.60,0.94), adjusting age, sex, and course of disease, the influence changed little. No DEFB4 gene CNV and body function (BASFI) associated.5.DEFB4_1: salt taste moderate patients were compared with salt flavored patients, gene * environmental interaction OR=0.51,95%CI (0) .28,0.94), P=0.032, other environmental factors did not find the interaction with the gene,.DEFB4_2 did not find the interaction between the copy number variation and the environment. Conclusion the high copy number of the DEFB4 gene (> 4) is a protective factor for the pathogenesis of AS, and is independent of age and sex; the increase of the copy number of the gene CNV is independent of age, sex and course of disease are serious to the disease. The degree of protection was increased, but the association between the increase of gene CNV and the physical function of the patient was not found, and the genetic environmental interaction analysis suggested that the dietary taste may affect the risk of AS with the variation of the copy number of the DEFB4 gene.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.23

【共引文献】