血清IL-10水平及其基因多态性在5岁以下喘息儿童中的研究
发布时间：2018-09-14 13:37
【摘要】：对5岁以下的儿童这一特殊群体来讲,能引起喘息的疾病种类原因颇多,如何鉴别是困扰儿科呼吸专业医生的难题。研究结果表明有约40%的支气管哮喘及反复喘息患儿第一次喘息发生在一岁以内。国内、国外的许多研究资料证明,早期、规范的对儿童哮喘进行干预及管理,将有效改善疾病预后、提高患者生活质量并降低社会医疗负担。目前有越来越多的研究试图找到一种确切的实验室指标,用以从小于5岁的喘息儿童中鉴别出真正的支气管哮喘病人,但目前这个问题尚未解决。 目的 本临床研究对IL-10基因转录启动子的单核苷酸多态性(single-nucleotide polymorphisms,SNPs)进行研究,同时观察外周血IL-10、IgE及EOS水平,了解反复喘息患儿发作时体内变化,尤其是结合哮喘预测指数的不同与正常儿童进行比较,以探讨IL-10与5岁以下儿童反复喘息发病机制之间的关系。 材料和方法 本实验设计有喘息组与对照组。选择2010年12月-2011年4月在郑州大学第三附属医院儿内科门诊就诊或收入病房治疗的喘息患儿共152例入选喘息组,喘息患儿均为中国河南省汉族儿童,均处于急性发作期,年龄小于或等于5岁。入选患儿在最近1年内至少发作4次喘息或最近半年内已至少发作2次喘息,排除早产儿、先天性心脏病儿童、气道发育畸形儿童、足月小样儿。根据患儿本人有医师诊断的变应性疾病(例如变应性鼻炎、湿疹、食物过敏、变应性皮炎)或患儿父母任一方有医师诊断的哮喘病史,作为特应质高危因素。根据研究对象是否具有高危因素将喘息患儿又分为2组,具有高危因素之一的患儿入选喘息Ⅰ组(n=78；男56例,女22例),不具有高危因素的患儿入选喘息Ⅱ组(n=74；男52例,女22例)。随机选择同时期本院儿童保健部健康体检儿童46例(n=46；男26例,女20例)入选健康对照组,对照组儿童均排除特应质高危因素。所有研究对象于近2周内均未患感染性疾病、未应用过激素类药物及其他免疫调节药物,所有入组儿童均为足月出生儿童,3组患儿年龄、性别进行比较,均无统计学差异(P0.05),入组前均告知其监护人并征得其知情同意。 采用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)方法检测IL-10启动子-592位点A/C基因多态性,比较三组研究对象IL-10启动子-592位点的基因型频率及等位基因频率。采用酶联免疫吸附实验法(ELISA)对3组患儿外周血IL-10及总IgE水平进行定量测定。采用伊红染色法计数外周血EOS。对三组患儿的外周血IL-10、总IgE及EOS计数水平的均值进行比较,并进一步分析有无相关性。 结果 1.三组中均可检测到CC、AC、AA3种基因型,并且三组均以杂合子AC型比率较高,野生型CC型比率次之,纯合突变型AA型比率较小。其中喘息Ⅰ组CC、AC、AA基因型频率为25.6%、59.0%、15.4%,A、C等位基因频率为44.9%、55.1%；喘息Ⅱ组CC、AC、AA基因型频率分别为21.6%、63.5%、14.9%,A、C等位基因频率为46.6%、53.4%；对照组CC、AC、AA基因型频率为37.0%、52.2%、10.8%,A、C等位基因频率为37%、63%。三组间比较各基因型频率及等位基因频率分布差异均无统计学意义(P值均0.05)。 2.喘息Ⅰ组外周血IL-10、总IgE水平及EOS计数水平均高于喘息Ⅱ组及对照组(P值均0.01),差异有统计学意义；喘息Ⅱ组外周血IL-10、总IgE水平均高于对照组(P值均0.01),差异有统计学意义,喘息Ⅱ组EOS计数水平与对照组比较,差异无统计学意义(P值0.05)。 3.相关性分析显示喘息Ⅰ组外周血IL-10水平与IgE水平呈正相关关系(r=0.815,P0.01),喘息Ⅱ组及对照组IL-10与EOS,IL-10与IgE均无相关。 结论 IL-10基因启动子-592位点基因多态性与5岁及其以下儿童反复喘息性疾病无相关性,但外周血IL-10水平与特应质及机体过敏因素存在高度相关,IL-10与具有特应质的5岁以下反复喘息儿童发病相关。
[Abstract]:For a special group of children under 5 years old, there are many causes of asthma, and how to identify them is a difficult problem for pediatric respiratory specialists. Fan's intervention and management of childhood asthma will effectively improve the prognosis of the disease, improve the quality of life and reduce the burden of social health care. Not solved.
objective
The single-nucleotide polymorphisms (SNPs) of IL-10 gene transcription promoter were studied in this clinical study. The levels of IL-10, IgE and EOS in peripheral blood were also observed. The changes in vivo in children with recurrent wheezing, especially in combination with different predictive indices of asthma, were compared with those in normal children. The following is the relationship between recurrent wheezing and the pathogenesis of children.
Materials and methods
A total of 152 children with asthma who were treated in the outpatient department of pediatric medicine or in the ward of the Third Affiliated Hospital of Zhengzhou University from December 2010 to April 2011 were enrolled in the asthma group. All the children were Han nationality children in Henan Province, China, who were in the acute attack stage and were younger than or equal to 5 years old. There have been at least four episodes of wheezing in the last year or at least two episodes of wheezing in the last six months, excluding premature infants, children with congenital heart disease, children with airway malformations, and full-term infants. History of asthma diagnosed by physicians as a high risk factor for atopic asthma. Children with asthma were divided into two groups according to whether the subjects had high risk factors. Children with one of the high risk factors were enrolled in asthma group I (n = 78; 56 males, 22 females), and children without high risk factors were enrolled in asthma group II (n = 74; 52 males, 22 females). 46 children (n = 46; 26 males, 20 females) were enrolled in the healthy control group. The control group excluded the high-risk factors of atopic substances. All the subjects had no infectious diseases in the past two weeks, and no hormone drugs or other immunomodulatory drugs were used. All the children in the study group were full-term children, and the three groups were full-term children. There was no significant difference in age and sex (P 0.05). The guardians were informed and informed consent was obtained before admission.
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the A/C gene polymorphism at the promoter-592 locus of IL-10. Genotype frequencies and allele frequencies at the promoter-592 locus of IL-10 were compared among the three groups. The levels of IL-10 and total IgE in peripheral blood of the three groups were measured by enzyme-linked immunosorbent assay (ELISA). The levels of IL-10, total IgE and EOS in peripheral blood of the three groups were compared, and the correlation was further analyzed.
Result
1. Three genotypes of CC, AC and AA were detected in all three groups, and the ratio of heterozygote AC genotype was higher in all three groups, followed by wild type CC genotype and homozygous mutant AA genotype. The allele frequencies of CC, AC and AA were 37.0%, 52.2%, 10.8%, 37%, 63.5%, 14.9%, 46.6% and 53.4% respectively, while those of control group were 37.0%, 52.2%, 10.8%, A and C allele frequencies were 37% and 63%, respectively. There was no significant difference among the three groups (P value was 0.05).
2. The levels of IL-10, total IgE and EOS in peripheral blood of asthmatic group I were higher than those of asthmatic group II and control group (P 0.01), and the difference was statistically significant. The levels of IL-10 and total IgE in peripheral blood of asthmatic group II were higher than those of control group (P 0.01), and the difference was statistically significant. (P value 0.05).
3. Correlation analysis showed that there was a positive correlation between IL-10 level and IgE level in peripheral blood of asthmatic group I (r=0.815, P 0.01). There was no correlation between IL-10 and EOS, IL-10 and IgE in asthmatic group II and control group.
conclusion
IL-10 promoter-592 gene polymorphism was not associated with recurrent wheezing disorders in children under 5 years old, but the levels of IL-10 in peripheral blood were highly correlated with atopy and allergic factors. IL-10 was associated with recurrent wheezing disorders in children under 5 years old with atopy.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.6
[Abstract]:For a special group of children under 5 years old, there are many causes of asthma, and how to identify them is a difficult problem for pediatric respiratory specialists. Fan's intervention and management of childhood asthma will effectively improve the prognosis of the disease, improve the quality of life and reduce the burden of social health care. Not solved.
objective
The single-nucleotide polymorphisms (SNPs) of IL-10 gene transcription promoter were studied in this clinical study. The levels of IL-10, IgE and EOS in peripheral blood were also observed. The changes in vivo in children with recurrent wheezing, especially in combination with different predictive indices of asthma, were compared with those in normal children. The following is the relationship between recurrent wheezing and the pathogenesis of children.
Materials and methods
A total of 152 children with asthma who were treated in the outpatient department of pediatric medicine or in the ward of the Third Affiliated Hospital of Zhengzhou University from December 2010 to April 2011 were enrolled in the asthma group. All the children were Han nationality children in Henan Province, China, who were in the acute attack stage and were younger than or equal to 5 years old. There have been at least four episodes of wheezing in the last year or at least two episodes of wheezing in the last six months, excluding premature infants, children with congenital heart disease, children with airway malformations, and full-term infants. History of asthma diagnosed by physicians as a high risk factor for atopic asthma. Children with asthma were divided into two groups according to whether the subjects had high risk factors. Children with one of the high risk factors were enrolled in asthma group I (n = 78; 56 males, 22 females), and children without high risk factors were enrolled in asthma group II (n = 74; 52 males, 22 females). 46 children (n = 46; 26 males, 20 females) were enrolled in the healthy control group. The control group excluded the high-risk factors of atopic substances. All the subjects had no infectious diseases in the past two weeks, and no hormone drugs or other immunomodulatory drugs were used. All the children in the study group were full-term children, and the three groups were full-term children. There was no significant difference in age and sex (P 0.05). The guardians were informed and informed consent was obtained before admission.
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the A/C gene polymorphism at the promoter-592 locus of IL-10. Genotype frequencies and allele frequencies at the promoter-592 locus of IL-10 were compared among the three groups. The levels of IL-10 and total IgE in peripheral blood of the three groups were measured by enzyme-linked immunosorbent assay (ELISA). The levels of IL-10, total IgE and EOS in peripheral blood of the three groups were compared, and the correlation was further analyzed.
Result
1. Three genotypes of CC, AC and AA were detected in all three groups, and the ratio of heterozygote AC genotype was higher in all three groups, followed by wild type CC genotype and homozygous mutant AA genotype. The allele frequencies of CC, AC and AA were 37.0%, 52.2%, 10.8%, 37%, 63.5%, 14.9%, 46.6% and 53.4% respectively, while those of control group were 37.0%, 52.2%, 10.8%, A and C allele frequencies were 37% and 63%, respectively. There was no significant difference among the three groups (P value was 0.05).
2. The levels of IL-10, total IgE and EOS in peripheral blood of asthmatic group I were higher than those of asthmatic group II and control group (P 0.01), and the difference was statistically significant. The levels of IL-10 and total IgE in peripheral blood of asthmatic group II were higher than those of control group (P 0.01), and the difference was statistically significant. (P value 0.05).
3. Correlation analysis showed that there was a positive correlation between IL-10 level and IgE level in peripheral blood of asthmatic group I (r=0.815, P 0.01). There was no correlation between IL-10 and EOS, IL-10 and IgE in asthmatic group II and control group.
conclusion
IL-10 promoter-592 gene polymorphism was not associated with recurrent wheezing disorders in children under 5 years old, but the levels of IL-10 in peripheral blood were highly correlated with atopy and allergic factors. IL-10 was associated with recurrent wheezing disorders in children under 5 years old with atopy.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.6
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