【摘要】：對(duì)5歲以下的兒童這一特殊群體來(lái)講,能引起喘息的疾病種類原因頗多,如何鑒別是困擾兒科呼吸專業(yè)醫(yī)生的難題。研究結(jié)果表明有約40%的支氣管哮喘及反復(fù)喘息患兒第一次喘息發(fā)生在一歲以內(nèi)。國(guó)內(nèi)、國(guó)外的許多研究資料證明,早期、規(guī)范的對(duì)兒童哮喘進(jìn)行干預(yù)及管理,將有效改善疾病預(yù)后、提高患者生活質(zhì)量并降低社會(huì)醫(yī)療負(fù)擔(dān)。目前有越來(lái)越多的研究試圖找到一種確切的實(shí)驗(yàn)室指標(biāo),用以從小于5歲的喘息兒童中鑒別出真正的支氣管哮喘病人,但目前這個(gè)問(wèn)題尚未解決。 目的 本臨床研究對(duì)IL-10基因轉(zhuǎn)錄啟動(dòng)子的單核苷酸多態(tài)性(single-nucleotide polymorphisms,SNPs)進(jìn)行研究,同時(shí)觀察外周血IL-10、IgE及EOS水平,了解反復(fù)喘息患兒發(fā)作時(shí)體內(nèi)變化,尤其是結(jié)合哮喘預(yù)測(cè)指數(shù)的不同與正常兒童進(jìn)行比較,以探討IL-10與5歲以下兒童反復(fù)喘息發(fā)病機(jī)制之間的關(guān)系。 材料和方法 本實(shí)驗(yàn)設(shè)計(jì)有喘息組與對(duì)照組。選擇2010年12月-2011年4月在鄭州大學(xué)第三附屬醫(yī)院兒內(nèi)科門診就診或收入病房治療的喘息患兒共152例入選喘息組,喘息患兒均為中國(guó)河南省漢族兒童,均處于急性發(fā)作期,年齡小于或等于5歲。入選患兒在最近1年內(nèi)至少發(fā)作4次喘息或最近半年內(nèi)已至少發(fā)作2次喘息,排除早產(chǎn)兒、先天性心臟病兒童、氣道發(fā)育畸形兒童、足月小樣兒。根據(jù)患兒本人有醫(yī)師診斷的變應(yīng)性疾病(例如變應(yīng)性鼻炎、濕疹、食物過(guò)敏、變應(yīng)性皮炎)或患兒父母任一方有醫(yī)師診斷的哮喘病史,作為特應(yīng)質(zhì)高危因素。根據(jù)研究對(duì)象是否具有高危因素將喘息患兒又分為2組,具有高危因素之一的患兒入選喘息Ⅰ組(n=78；男56例,女22例),不具有高危因素的患兒入選喘息Ⅱ組(n=74；男52例,女22例)。隨機(jī)選擇同時(shí)期本院兒童保健部健康體檢兒童46例(n=46；男26例,女20例)入選健康對(duì)照組,對(duì)照組兒童均排除特應(yīng)質(zhì)高危因素。所有研究對(duì)象于近2周內(nèi)均未患感染性疾病、未應(yīng)用過(guò)激素類藥物及其他免疫調(diào)節(jié)藥物,所有入組兒童均為足月出生兒童,3組患兒年齡、性別進(jìn)行比較,均無(wú)統(tǒng)計(jì)學(xué)差異(P0.05),入組前均告知其監(jiān)護(hù)人并征得其知情同意。 采用聚合酶鏈?zhǔn)椒磻?yīng)-限制性片段長(zhǎng)度多態(tài)性(PCR-RFLP)方法檢測(cè)IL-10啟動(dòng)子-592位點(diǎn)A/C基因多態(tài)性,比較三組研究對(duì)象IL-10啟動(dòng)子-592位點(diǎn)的基因型頻率及等位基因頻率。采用酶聯(lián)免疫吸附實(shí)驗(yàn)法(ELISA)對(duì)3組患兒外周血IL-10及總IgE水平進(jìn)行定量測(cè)定。采用伊紅染色法計(jì)數(shù)外周血EOS。對(duì)三組患兒的外周血IL-10、總IgE及EOS計(jì)數(shù)水平的均值進(jìn)行比較,并進(jìn)一步分析有無(wú)相關(guān)性。 結(jié)果 1.三組中均可檢測(cè)到CC、AC、AA3種基因型,并且三組均以雜合子AC型比率較高,野生型CC型比率次之,純合突變型AA型比率較小。其中喘息Ⅰ組CC、AC、AA基因型頻率為25.6%、59.0%、15.4%,A、C等位基因頻率為44.9%、55.1%；喘息Ⅱ組CC、AC、AA基因型頻率分別為21.6%、63.5%、14.9%,A、C等位基因頻率為46.6%、53.4%；對(duì)照組CC、AC、AA基因型頻率為37.0%、52.2%、10.8%,A、C等位基因頻率為37%、63%。三組間比較各基因型頻率及等位基因頻率分布差異均無(wú)統(tǒng)計(jì)學(xué)意義(P值均0.05)。 2.喘息Ⅰ組外周血IL-10、總IgE水平及EOS計(jì)數(shù)水平均高于喘息Ⅱ組及對(duì)照組(P值均0.01),差異有統(tǒng)計(jì)學(xué)意義；喘息Ⅱ組外周血IL-10、總IgE水平均高于對(duì)照組(P值均0.01),差異有統(tǒng)計(jì)學(xué)意義,喘息Ⅱ組EOS計(jì)數(shù)水平與對(duì)照組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P值0.05)。 3.相關(guān)性分析顯示喘息Ⅰ組外周血IL-10水平與IgE水平呈正相關(guān)關(guān)系(r=0.815,P0.01),喘息Ⅱ組及對(duì)照組IL-10與EOS,IL-10與IgE均無(wú)相關(guān)。 結(jié)論 IL-10基因啟動(dòng)子-592位點(diǎn)基因多態(tài)性與5歲及其以下兒童反復(fù)喘息性疾病無(wú)相關(guān)性,但外周血IL-10水平與特應(yīng)質(zhì)及機(jī)體過(guò)敏因素存在高度相關(guān),IL-10與具有特應(yīng)質(zhì)的5歲以下反復(fù)喘息兒童發(fā)病相關(guān)。
[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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R725.6

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