哮喘易感基因HLA-DQ、ORMDL3单核苷酸多态性在婴幼儿喘息中的研究

发布时间：2018-06-23 02:23

本文选题：婴幼儿喘息 + 哮喘　；参考：《山东中医药大学》2017年硕士论文

【摘要】：目的:探讨哮喘易感基因HLA-DQA1、HLA-DQA2、ORMDL3单核苷酸多态性在婴幼儿喘息中的分布规律,为哮喘的早期诊断提供理论依据。方法:收集2015年10月至2016年11月在山东省立医院小儿呼吸科住院及门诊喘息患儿150例,其中API阳性组喘息患儿80例,API阴性组喘息患儿70例。无菌采集静脉血3ml,采用天根核酸提取仪进行基因组DNA提取,并通过96.96集成流体通路(IFC)Juno系统对哮喘易感基因HLA-DQA1单核苷酸多态性(SNP)位点rs9272346、HLA-DQA2 SNP位点rs7773955以及ORMDL3 SNPs位点rs4794820、rs7216389基因型进行检测,比较API阳性组和API阴性组喘息患儿基因型的分布差异,分析喘息患儿的潮气肺功能、呼出气一氧化氮(FENO)浓度、外周血嗜酸粒细胞百分数(EOS%)、血清免疫球蛋白E(Ig E)与基因型分布的相关性,并通过广义多因子降维法(GMDR)探讨上述哮喘易感基因间的交互作用。结果:1.rs9272346位点在API阳性组中以杂合子AG基因型的频率最高,纯合子AA基因型的频率最低,GG基因型的频率介于AG与AA之间;在API阴性组中以杂合子AG基因型的频率最高,纯合子GG基因型的频率最低,AA基因型的频率介于AG与GG之间。两组间基因型分布无统计学差异(P0.05)。2.rs7773955位点在API阳性组与API阴性组中均以纯合子CC基因型的频率最高,TT基因型的频率最低,杂合子TC基因型的频率介于CC与TT之间。基因型CC在API阳性组的分布频率明显高于阴性组,差异有统计学意义(χ2=6.561,P=0.010,OR=2.345,95%CI=1.215~4.524),TT基因型在API阴性组的分布频率明显高于阳性组,差异有统计学意义(χ2=6.381,P=0.012,OR=0.267,95%CI=0.091~0.784)。3.rs4794820位点在API阳性组中以杂合子AG基因型的频率最高,纯合子AA基因型的频率最低,GG基因型的频率介于AG与AA之间;在API阴性组中以纯合子GG基因型的频率最高,AA基因型的频率最低,杂合子AG基因型的频率GG与AA之间。AG基因型在API阳性组的分布频率明显高于阴性组,差异有统计学意义(χ2=11.219,P=0.001,OR=3.107,95%CI=1.586~6.090),GG基因型在API阴性组的分布频率明显高于阳性组,差异有统计学意义(χ2=12.411,P=0.000,OR=0.303,95%CI=0.154~0.595)。4.rs7216389位点基因型分布频率在API阳性组中以杂合子TC为最高,纯合子TT其次,CC最低;在API阴性组中以纯合子TT为最高,杂合子TC其次,CC最低。TC基因型在API阳性组的分布频率明显高于阴性组,差异有统计学意义(χ2=8.244,P=0.004,OR=2.627,95%CI=1.349~5.115),TT基因型在API阴性组的分布频率明显高于阳性组,差异有统计学意义(χ2=10.193,P=0.001,OR=0.341,95%CI=0.175~0.666)。5.GMDR分析显示有统计学意义的最佳交互模型包括多态性位点rs9272346、rs7773955、rs4794820、rs72163895,该模型的检测样本准确度为0.6946,交叉验证一致性的结果为10/10,应用Logistic分析显示上述四个位点的联合作用模式为发生API阳性婴幼儿喘息的独立危险因素(OR=1.026,95%CI:1.006-1.046,P=0.011)。6.在API阳性组中,哮喘易感基因ORMDL3SNPs位点rs4794820以及rs7216389同时表达AG/TC基因型患儿(44例)的TPTEF/TE和VPEF/VE值均低于同时表达GG/TT基因型的患儿(23例)(分别为14.55±4.83和19.91±4.17、18.85±4.26和25.20±7.06);FENO浓度、EOS%、Ig E水平则均高于同时表达GG/TT基因型的患儿(分别为25.02±8.77ppb和18.39±6.56ppb、7.16±2.62和5.50±1.34、366.73±275.53 IU/m L和166.83±62.87IU/m L),差异均有统计学意义(|t|=4.727、3.976、3.484、3.409、4.589,P均0.01)。7.API阳性组患儿的达峰时间比(TPTEF/TE)、达峰容积比(VPEF/VE)均低于API阴性组(分别为16.87±5.31和20.12±5.23、20.87±5.92和25.56±6.77),FENO浓度高于阴性组(22.44±9.77ppb和13.23±7.90ppb),差异均有统计学意义(|t|=3.776、4.490、6.377,P均0.01)。8.在API阴性组与阳性组中,FENO与TPTEF/TE及VPEF/VE均无相关性(P均0.05)。结论:1.哮喘易感基因HLA-DQA2 SNP位点rs7773955 CC基因型、ORMDL3 SNPs位点rs4794820 AG基因型以及rs7216389 TC基因型在API阳性组中分布频率明显增高,为婴幼儿喘息发病的高危基因型;2.哮喘易感基因ORMDL3 SNPs位点rs4794820以及rs7216389同时表达AG、TC基因型的API阳性喘息患儿的肺功能损害程度及气道炎症水平显著高于同时表达GG、TT基因型者,为婴幼儿重症喘息的高危人群;3.四个SNPs位点rs9272346、rs7773955、rs4794820、rs72163895之间存在正交互作用,协同增强了API阳性婴幼儿喘息的发病风险,提高了喘息患儿发展为持续性哮喘的危险性。
[Abstract]:Objective: To investigate the distribution of single nucleotide polymorphisms of HLA-DQA1, HLA-DQA2, ORMDL3 in asthma in infants and infants, and to provide a theoretical basis for early diagnosis of asthma. Methods: 150 cases of asthmatic children in the pediatric department of respiration in Shangdong Province-owned Hospital from October 2015 to November 2016 were collected, and 80 of the children were wheezing in the positive group of the Shangdong Province-owned Hospital. In the API negative group, 70 children with wheezing were collected aseptic venous blood 3ml, genomic DNA was extracted by RNA extraction instrument, and the HLA-DQA1 single nucleotide polymorphisms (SNP) rs9272346, HLA-DQA2 SNP locus rs7773955 and ORMDL3 sites were used by the 96.96 integrated fluid pathway (IFC) Juno system. Genotypes were detected to compare the distribution difference between the API positive group and the API negative group, and analyze the tidal gas and lung function, the concentration of exhaled nitric oxide (FENO), the percentage of eosinophils (EOS%) in peripheral blood (EOS%), the correlation of serum immunoglobulin E (Ig E) and the genotype distribution, and through the generalized multifactor reduction method (GMD). R) to explore the interaction between the susceptibility genes of the above asthma. Results: the frequency of the heterozygote AG genotype in the API positive group is the highest, the frequency of the homozygote AA genotype is the lowest, the frequency of the GG genotype is between AG and AA, and the frequency of the heterozygote AG based genotype is the highest in the API negative group and the frequency of the homozygote GG genotype is the most. The frequency of AA genotype was between AG and GG. There was no statistical difference between the two groups (P0.05), the frequency of CC genotype of YISHION zygote in API positive group and API negative group was the highest, the frequency of TT genotype was the lowest, and the frequency of the heterozygote TC genotypes was between CC and TT. The frequency was significantly higher than that of the negative group, the difference was statistically significant (x 2=6.561, P=0.010, OR=2.345,95%CI=1.215~4.524), the distribution frequency of TT genotype in API negative group was significantly higher than that of the positive group, and the difference was statistically significant (x 2=6.381, P=0.012, OR=0.267,95%CI=0.091~0.784).3.rs4794820 locus in API positive group with heterozygote AG genotype frequency The frequency of the homozygote AA genotype is the lowest, the frequency of the GG genotype is between AG and AA, and the frequency of the GG genotype of YISHION zygote is the highest in the API negative group, the frequency of the AA genotype is the lowest. The frequency of the frequency GG of the heterozygote AG genotype and the.AG genotype between AA is significantly higher than that of the negative group, and the difference is statistically significant. The distribution frequency of GG genotype in API negative group was significantly higher than that of the positive group (x 2=11.219, P=0.001, OR=3.107,95%CI=1.586~6.090), and the difference was statistically significant (x 2=12.411, P=0.000, OR=0.303,95%CI=0.154~0.595) and the frequency of genotype distribution in the API positive group was the highest in the API positive group, and the homozygote TT was the next and the lowest. In the negative group, the TT of YISHION zygote was the highest and the heterozygote TC was second. The distribution frequency of the lowest.TC genotype in the API positive group was significantly higher than that in the negative group. The difference was statistically significant (x 2=8.244, P=0.004, OR=2.627,95%CI=1.349~5.115), and the frequency of TT genotype in the API negative group was significantly higher than that of the positive group (x 2=10.193, P=0). .001, OR=0.341,95%CI=0.175~0.666).5.GMDR analysis showed that the best interactive model of statistical significance included polymorphic loci rs9272346, rs7773955, rs4794820, rs72163895, the accuracy of the sample was 0.6946, the consistency of cross validation was 10/10, and the combination of the above four sites should be displayed with Logistic analysis. The independent risk factor (OR=1.026,95%CI:1.006-1.046, P=0.011) for API positive infants (OR=1.026,95%CI:1.006-1.046, P=0.011) was in the API positive group, the ORMDL3SNPs locus rs4794820 of the asthma susceptibility gene and the rs7216389 in the AG/TC genotype children (44 cases) were lower than those of the children (23 cases) that expressed the GG/TT genotypes (14.55 respectively). The concentration of FENO, EOS%, Ig E were higher than those of GG/TT genotypes (25.02 + 8.77ppb and 18.39 + 6.56ppb, 7.16 + 2.62 and 5.50 + 1.34366.73 + 275.53 IU/m L and 166.83 + 62.87IU/m). The difference was statistically significant. 4.83 .01) the peak time ratio of children in.7.API positive group (TPTEF/TE) and peak volume ratio (VPEF/VE) were lower than that of API negative group (16.87 + 5.31 and 20.12 + 5.23,20.87 + 5.92 and 25.56 + 6.77 respectively). The concentration of FENO was higher than that of negative group (22.44 + 9.77ppb and 13.23 + 7.90ppb), and the difference was statistically significant (|t|=3.776,4.490,6.377, P are 0.01). In the positive group, there was no correlation between FENO and TPTEF/TE and VPEF/VE (P 0.05). Conclusion: 1. the rs7773955 CC genotypes of the HLA-DQA2 SNP loci of the asthma susceptible genes, the ORMDL3 SNPs locus rs4794820 AG genotypes and the genotype in the positive group are significantly higher, which is the high-risk genotype of infant wheezing; 2. asthma susceptibility. The gene ORMDL3 SNPs locus rs4794820 and rs7216389 simultaneously expressed AG, and the degree of lung function damage and airway inflammation in API positive asthmatic children with TC genotypes were significantly higher than that of simultaneous expression of GG. The TT genotype was the high risk population of infants with severe wheezing; 3. four SNPs loci were rs9272346, rs7773955, Interaction enhances the risk of wheezing in API positive infants, and increases the risk of developing asthmatic children to persistent asthma.
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R725.6

【参考文献】