ACE, CD14, IL4基因多型性與台灣過敏疾病關聯性之研究

The Association Study of ACE, CD14, IL-4 Gene Polymorphism and Atopic Disease in Taiwan

顧明修(Min-Sho Ku)

指導教授 ： 黎慶 ； 呂克桓 ； 王志堯 ； 吳宗遠

中山醫學大學/醫學院/醫學研究所/碩士(2005年)

https://doi.org/10.6834/CSMU.2005.00058

摘要

過敏疾病包括過敏性鼻炎及氣喘，在台灣及全世界都是很常見的疾病。一般認為此病主要是環境及遺傳因素交互作用而造成的。近年來，關於遺傳因素的研究，包括位置選殖法 (positional cloning)，候選基因 (candidate gene) 的鑑定及候選基因之基因多型性 (candidate gene polymorphism) 已陸續被發表。但大部分都是外國的研究。因此我們的研究目的為探討候選基因之基因多型性和台灣人過敏疾病的關係。
血管張力素轉化酶 (ACE) 在第二型血管收縮素 (angiotensin II)形成及將tachykinin, bradykinin去活化上扮演ㄧ定的角色; 第二型血管收縮素為強力的氣管收縮素，tachykinin, bradykinin則為氣管發炎的媒介。在國外有些研究指出，ACE基因intron 16 插入/刪除基因多型性 (I/D polymorphism) 和過敏疾病的發生有關。CD14 為細菌脂多醣 (lipopolysaccharide) 的接受器，和未活化T細胞 (naive T cell) 轉換為第ㄧ型或第二型T細胞 (Th1 or Th2) 有關。國外有些研究指出CD14基因啟動子 (promoter) -159位置C/T單一核甘酸多型性 (single nucleotide polymorphism, SNP) 和過敏疾病有關。IL-4 和IgE 形成及其他過敏機制有關，國外已有報導指出IL-4 基因intron-2 中70個鹼基對重複多型性和血清IL-4濃度有關，但此基因多型性和過敏疾病的關係至今仍無相關報導。我們的研究將探討上述三個基因多型性和台灣人過敏疾病的關係。
我們隨機選擇108名兒童為過敏兒童組，其中54人有過敏性鼻炎症狀，54人有過敏性鼻炎併發氣喘症狀；另選擇38個過敏疾病家族為過敏家族組 (47個成人，49個兒童)。接著選擇96個健康兒童為對照組。 ACE, IL-4 基因型藉由DNA聚合酶連鎖反應 (Polymerase Chain Reaction , PCR) 得知，CD14 基因型藉由PCR 及限制脢片段多型性 (Restriction fragment length polymorphism) 的方法得知。此外過敏症狀兒童利用肺功能機評估肺功\\\\能，血清總IgE值及台灣常見6種抗原特異性IgE值由Pharmacia cap system測得。血清嗜酸性球數量亦加以測之。接著我們比較實驗組，對照組間及不同情況下各基因型和各對偶基因 (allele) 比例的差別。IgE值，嗜酸性球數量，肺功能和各基因型的關係亦加以評估。
在ACE 基因多型性方面，“過敏性鼻炎併有氣喘”之兒童DD基因型出現的比例比控制組高( p = 0.008)，但此比例在“只有過敏性鼻炎”之兒童和控制組之間並無有意義的差別( p = 0.374)。我們同時發現有氣喘症狀之兒童，D對偶基因出現的比例在台灣人是35%，此比例小於中國的研究 (51%)。至於ACE基因型和血清IgE值，嗜酸性球數及 肺功能則沒有相關性。
　　在CD14，IL-4基因多型性方面，其基因型分配在實驗組及對照組並無差別，且和血清IgE值，嗜酸性血球數，肺功能間沒有相關性。在過敏家族，“有過敏症狀成員” 和“無過敏症狀成員”的三種基因型比例亦無差別。
由我們的研究得到的結論是，ACE 基因I/D多型性中的DD基因型和“過敏性鼻炎併有氣喘” 之症狀有關，和“只有過敏性鼻炎”之症狀無關。ACE 基因I/D多型性和血清IgE值，嗜酸性球數，肺功能間沒有相關性。CD14 基因C/T多型性，IL-4 intron 2 70鹼基對重複多型性和台灣人過敏疾病無關。

關鍵字

ACE ； CD14 ； IL4 ； 基因多型性 ； 台灣 ； 過敏疾病

並列摘要

Atopic disease including allergic rhinitis and asthma, are common in the world and Taiwan. The disease are genally considered to be caused by interaction of genetic and environmental factors. In recent years, studies about genetic factor, including positional cloning, candidate gene studies and polymorphisn of candidate genes are published. But most of the study are reported by other countries. So the aim of our study was to investigate the relationship of candidate genes polymorphism and atopic disease in Taiwan.
Angiotensin-converting enzyme (ACE) play a key role in the metabolism of angiotensin II and inactivation of bradykinins and tachykinins, which are potent bronchoconstrictors and mediators of airway inflammation seperatively. An insertion-deletion (I/D) polymorphism on intron 16 of ACE gene has been reported to be associated with atopic disease from other countries. CD14, a receptor of lipopolysaccharides, play a role in the polarization of naïve T cell to Th1 or Th2. C -159T single nucleotide polymorphism on CD14 gene promoter region has been reported to be associated with atopic disease from other countries. IL-4 is related to IgE production and other allergic reaction. IL-4 gene intron-2 70bp repeats polymorphism has been shown to be associated with serum IL-4 level, but it’s relationship to atopic disease are not reported now. Our study will investigate the relationship of the three genetic polymorphism and atopic disease in Taiwan.
We randomly select 108 children, 54 with allergic rhinitis symptom, 54 with allergic rhinitis combined asthma, as atopic children group; 38 atopic disease families including 47 adults and 49 children, as atopic disease group. Then select 96 healthy children as control group. ACE and IL-4 genotype were determined by PCR. CD14 genotype was determined by PCR and restriction analysis by Ava II. Lung function in atopic disease children was assessed by spirometry. Pharmacia cap system were performed to measure serum total IgE levels and and six commom allergen specific IgE levels. Serum eosinophil count also checked. Then we investigate the frequencies of the genotypes and alleles between study group and control group, and in different conditions. The relationship of genotype and IgE, eosinophil, lung function data also evaluated.
In ACE gene polymorphism, the frequency of DD genotype was hgher in “allergic rhinitis combined asthma” children than control children ( p = 0.008). But there was no difference in the frequency of DD genotype between “only allergic rhinitis children” and control children ( p = 0.374).
We also found that in Taiwaness asthma children, the rate of D allele are lower (35%) than that in Chinese asthma children (51%). Total IgE level, eosinophil count, lung function did not differe between ACE genotypes.
In CD14 and IL-4 gene polymorphism, the genotype distribution was not different between atopic children group and control group. Total IgE level, eosinophil count, lung function still did not differe between genotypes. In atopic disease family, the genotype distribution of the three genetic polymorphism was not different between “with atopic symptom members” and “without atopic symptom members”.
In conclusion, the result suggested that DD allele of ACE genotype was significantly associated with “allergic rhinitis combined asthma children”, but not “allergic rhinitis only children”. ACE I/D polymorphism is not related to lung function, serum total IgE level and eosinophil count. Both CD14 C/T polymorphism and IL-4 intron 2 70bp repeats polymorphism are not relate to atopic disease in Taiwaness.

並列關鍵字

ACE ； CD14 ； IL-4 ； Gene ； Polymorphism ； Atopic Disease ； Taiwan

參考文獻

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