哮喘患者的蛋白质微阵列分析
2009/05/18
微阵列技术为了解哮喘患者整个的基因和蛋白质表达序列提供了一种新方法。为了识别在哮喘患者的气道中产生的新的因子，Hyo-Bin Kim等人对有支气管哮喘的患者的痰标本应用了基膜人细胞因子微阵列技术（membrane-based human cytokine microarray technology）。
他们从28例支气管哮喘患者（BA）、20例无哮喘症状但有遗传性过敏症的病例(AC)，和38例无哮喘症状无遗传性过敏症的健康对照人群（NC）中取得诱导痰，并随机选取9例BA，3例AC，6例NC的诱导痰，取上层痰液进行常规人细胞因子阵列分析（custom human cytokine array），同时分析到了79种特异的细胞因子。
并通过ELISAs 法检测到了调节生长肿瘤基因(GRO)-α, 嗜酸细胞活化趋化因子-2, 以及肺调节活化趋化因子(PARC)/CCL18 水平，应用放射免疫测定出嗜酸粒细胞衍生神经毒素(EDN)。微阵列技术发现BA组较AC和NC组中的GRO-α, 嗜酸细胞活化趋化因子-2, 和PARC的信号强度均明显增强(p分别为0.036、 0.042和0.033)。ELISA 法检测BA组较AC和NC组中的PARC蛋白水平有明显增高(p < 0.0001)，且PARC水平与痰嗜酸细胞百分比(r = 0.570, p < 0.0001)、EDN—正常T细胞表达和分泌细胞因子水平(r = 0.440, p < 0.001)、白介素-4(r = 0.415, p < 0.01)、干拢素-γ (r = 0.491, p < 0.001)明显相关。通过无偏差无干扰的检测方法，PARC，作为一种趋化因子在哮喘患者的痰标本中被检测出来。
该研究提出PARC可能在哮喘的气道嗜酸性炎症反应中起着重要作用。
（于娜 中国医科大学附属第一医院呼吸科 110001 摘译）
（Chest February 2009 135:295-302; doi:10.1378/chest.08-0962）
（Chest February 2009 135:295-302; doi:10.1378/chest.08-0962）
Protein Microarray Analysis in Patients With Asthma Elevation of the Chemokine PARC/CCL18 in Sputum
Keywords: asthma 、cytokine microarray analysis 、eosinophilic inflammation 、pulmonary and activation-regulated chemokine/CCL18
Hyo-Bin Kim, MD, PhD†,
Chang-Keun Kim, MD, PhD†,
Koji Iijima, PhD,
Takao Kobayashi, PhD and
Hirohito Kita, MD*
Abstract
Background: Microarray technology offers a new opportunity to gain insight into global gene and protein expression profiles in asthma. To identify novel factors produced in the asthmatic airway, we analyzed sputum samples by using a
membrane-based human cytokine microarray technology in patients with bronchial asthma (BA).
Methods: Induced sputum was obtained from 28 BA subjects, 20 nonasthmatic atopic control (AC) subjects, and 38 nonasthmatic nonatopic normal control (NC) subjects. The microarray samples of subjects were randomly selected from nine BA subjects, three AC subjects, and six NC subjects. Sputum supernatants were analyzed using a custom human cytokine array (RayBio Custom Human Cytokine Array; RayBiotech; Norcross, GA) designed to analyze 79 specific cytokines simultaneously. The levels of growth-regulated oncogene (GRO)-α, eotaxin-2, and pulmonary and activation-regulated chemokine (PARC)/CCL18 were measured by sandwich enzyme-linked immunosorbent assays (ELISAs), and eosinophil-derived neurotoxin (EDN) was measured by radioimmunoassay.
Results: By microarray, the signal intensities for GRO-α, eotaxin-2, and PARC were significantly higher in BA subjects than in AC and NC subjects (p = 0.036, p = 0.042, and p = 0.033, respectively). By ELISA, the sputum PARC protein levels were significantly higher in BA subjects than in AC and NC subjects (p < 0.0001). Furthermore, PARC levels correlated significantly with sputum eosinophil percentages (r = 0.570, p < 0.0001) and the levels of EDN (r = 0.633, p < 0.0001), the regulated upon activation, normal T cell expressed and secreted cytokine (r = 0.440, p < 0.001), interleukin-4 (r = 0.415, p < 0.01), and interferon-γ (r = 0.491, p < 0.001).
Conclusions: By a nonbiased screening approach, a chemokine, PARC, is elevated in sputum specimens from patients with asthma. PARC may play important roles in development of airway eosinophilic inflammation in asthma.
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哮喘患者吸入特异性变应原对血清脂肪连接素（Adiponectin）的影响
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穿心莲内酯通过抑制NF-κB途径抑制哮喘发生
