HLA-DQA1基因多态性与儿童支气管哮喘的相关性及血清IL-4、IFN-γ水平关系的研究

发布时间：2018-01-25 04:45

本文关键词： 支气管哮喘 HLA-DQA1基因干扰素-γ 白介素-4　出处：《泸州医学院》2013年硕士论文　论文类型：学位论文

【摘要】：目的：探讨人类白细胞抗原HLA-DQA1基因多态性与儿童支气管哮喘的关联性，寻找与哮喘发作有关的易感基因和（或）抑制基因；观察哮喘组的血清IL-4、IFN-γ水平；比较含有及不含有哮喘相关基因的哮喘患者血清中IL-4、IFN-γ的浓度，分析是否有差异，为进一步研究哮喘发病机制、预防哮喘发作及寻找基因水平上治疗哮喘的方法提供相应的理论依据。方法：1.标本采集：哮喘组：选取急性发作期哮喘患儿共45例为哮喘组。对照组：收集与哮喘组年龄相仿的正常健康儿童60例为对照组。2.检测方法及指标：采用聚合酶链反应—序列特异性引物(PCR-SSP)方法测定哮喘组、对照组的HLA-DQAl等位基因，并计算两组中各等位基因的基因频率（直接计算法：基因频率=阳性基因数/样本数×2），两组间进行比较；采用双抗体夹心酶联免疫吸附法（ELISA）测定哮喘组、对照组中血清IL-4、IFN-γ水平浓度，并对结果进行统计学处理。3.统计学处理：运用SPPS-17.0软件系统分析，哮喘组与对照组各等位基因的基因频率比较采用χ2检验，P0.05认为有统计学意义，并计算优势比（OR）及95％置信区间。测定的血清IL-4、IFN-γ水平浓度，，经SPPS-17.0软件系统分析后结果用均数±标准差（±s）表示，两两比较采用LSD-t检验，相关分析采用Pearson相关性分析，P0.05认为有统计学意义。结果：（1）哮喘组与对照组之间HLA-DQA1等位基因频率比较：HLA-DQA1*0301在哮喘组的基因频率（20.00%）明显高于对照组（8.33%），χ2=7.159，P=0.007，有统计学意义，OR为3.333（95%置信区间为1.35-8.229）。HLA-DQA1*0104在哮喘组的基因频率（27.77%）明显低于对照组（38.33%），χ2值为5.234，P=0.022，差异有统计学意义，OR为0.380，95%置信区间为（0.164-0.880）。（2）哮喘组和对照组外周血IL-4、INF-γ水平比较：哮喘组中血清IL-4水平(116.45±20.45)明显高于对照组(90.34±13.43),差异有统计学意义（P0.05）；哮喘组血清IFN-γ水平(65.34±12.34)明显低于对照组(84.23±13.42)，差异有统计学意义（P0.05）；（3）哮喘组IFN-γ与IL-4的相关性分析：哮喘急性发作期血清IFN-γ与IL-4水平呈负相关（r=-0.356，P0.05）。（4）HLA-DQA1*0301阳性哮喘患者（18例）与HLA-DQA1*0301阴性哮喘患者（27例）IFN-γ（水平分别为67.12±13.24、69.13±12.16）、IL-4（水平分别为114.23±23.45、113.21±20.34）比较，差异无统计学意义（P0.05）。结论：1.HLA-DQA1*0301基因可能为儿童支气管哮喘的易感基因；HLA-DQA1*0104基因可能为儿童哮喘的免疫抑制基因。2.哮喘组中IFN-γ较对照组明显下降，IL-4则明显升高，哮喘组中两者呈负相关，表明Th1/Th2免疫失衡仍可能是哮喘发作的重要因素。3.存在与不存在易感基因的哮喘患者血清IFN-γ、IL-4水平比较无显著差异，表明HLA-DQA1*0301可能仅某种特应性变应原所致的哮喘有关，亦表明哮喘发作与否不是由某个等位基因决定，其发作可能为多基因共同作用的结果。
[Abstract]:Objective: to investigate the association between polymorphisms of human leukocyte antigen (HLA-DQA1) gene and bronchial asthma in children, and to search for susceptibility genes and / or suppressor genes related to asthma attack. The levels of IL-4 and IFN- 纬 in serum of asthmatic group were observed. To compare the levels of IL-4 and IFN- 纬 in the serum of asthmatic patients with and without asthmatic related genes, and to analyze whether there are differences in the levels of IL-4 and IFN- 纬, so as to further study the pathogenesis of asthma. Methods 1. Specimen collection: asthma group: 45 children with acute asthma were selected as asthma group. 60 healthy children of the same age as asthma group were collected as control group. 2. Detection methods and indicators: polymerase chain reaction-sequence specific primers (PCR) were used. The asthma group was determined by PCR-SSP. The HLA-DQAl allele of the control group and the gene frequency of each allele in the two groups were calculated (direct calculation method: gene frequency = number of positive gene / sample number 脳 2G), the comparison was made between the two groups. The serum levels of IL-4 and IFN- 纬 in asthma group and control group were determined by double antibody sandwich enzyme linked immunosorbent assay (Elisa). Statistical analysis: using SPPS-17.0 software, the allele frequencies of asthma group and control group were compared by 蠂 2 test. P0.05 thought there was statistical significance, and calculated the odds ratio (OR) and 95% confidence interval. The serum IL-4 level of IFN- 纬 was measured. The results were expressed as mean 卤standard deviation (卤s) after systematic analysis by SPPS-17.0 software. LSD-t test was used for pairwise comparison and Pearson correlation analysis was used for correlation analysis. Results the allele frequency of HLA-DQA1 between the asthma group and the control group was higher than that of the control group (P < 0.05). (20.00) was significantly higher than the control group (8.33). 蠂 2 = 7.159%, P = 0.007, with statistical significance. The OR was 3.333 / 95% CI = 1.35-8.229.HLA-DQA1 / 0104 gene frequency in asthma group was significantly lower than that in control group (P < 0.05). 38.33). 蠂 ~ 2 value was 5.234 (P = 0.022), the difference was statistically significant (OR = 0.380). The 95% confidence interval was 0.164-0.880) IL-4 in peripheral blood of asthmatic group and control group. Comparison of INF- 纬 levels: serum IL-4 levels in asthma group were significantly higher than those in control group (116.45 卤20.45) and 90.34 卤13.43). The difference was statistically significant (P 0.05). The level of IFN- 纬 in asthmatic group (65.34 卤12.34) was significantly lower than that in control group (84.23 卤13.42), and the difference was statistically significant (P 0.05). (3) correlation analysis between IFN- 纬 and IL-4 in asthma group: serum IFN- 纬 and IL-4 level were negatively correlated with IL-4 in acute episode of asthma. There were 18 patients with HLA-DQA1-0301 positive asthma and 27 patients with HLA-DQA1*0301 negative asthma. The levels were 67.12 卤13.24 respectively. Comparison of IL-4 levels (114.23 卤23.45, 113.21 卤20.34). Conclusion: 1. HLA-DQA1 / 0301 gene may be a susceptible gene for bronchial asthma in children. HLA-DQA1*0104 gene may be the immunosuppressive gene of childhood asthma. IFN- 纬 in asthma group was significantly lower than that in control group and IL-4 was significantly increased. There was a negative correlation between the two in asthma group, indicating that Th1/Th2 immune imbalance may still be an important factor of asthma attack. 3. The presence and absence of susceptibility gene in the serum IFN- 纬 of asthma patients. There was no significant difference in IL-4 levels, suggesting that HLA-DQA1*0301 might be related only to asthma caused by an atopic allergen, and that asthma attack was not determined by an allele. The attack may be the result of multiple gene interaction.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R725.6

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