O'Driscoll B R, Cromwell O, Kay A B
Clin Exp Immunol. 1984 Feb;55(2):397-404.
Sputum samples from patients with bronchial asthma, chronic bronchitis and cystic fibrosis were examined for the presence of leukotrienes B4, C4 and D4. Following ethanol extraction and purification on Amberlite XAD-8, leukotrienes were identified by high pressure liquid chromatography (HPLC) using the appropriate markers. Fractions from HLPC were also tested for biological activity using both the Boyden chemotaxis assay and FPL 55712 inhibitable contraction of the isolated guinea-pig ileum. LTB4 was detected in the HPLC fractionated sputa from bronchial asthma (seven of seven), chronic bronchitis (four of four) and cystic fibrosis (four of four). In contrast, bioassay on the guinea-pig ileum failed to detect LTC4 or LTD4 in 17 asthmatic sputa, although they were detected in one of five bronchitics and 16 of 25 patients with cystic fibrosis. The activity in eight of these cystic fibrosis sputa was further characterized by HPLC and shown to be LTC4 and/or LTD4. Sputum from 11 of 17 asthmatics, four of 25 patients with cystic fibrosis and two of five bronchitics contained an anaphylatoxin like substance. The majority of sputum samples containing LTB4 also possessed an activity with physical and biological characteristics of the 5(S), 12(S), 6-trans LTB4 isomer. These studies indicate that lipoxygenase products of arachidonic acid metabolism are present in the sputum in various forms of obstructive airways disease. The failure to detect the 'SRS-A' leukotrienes in sputum from bronchial asthma may be attributable to either losses during extraction, the insensitivity of the assay procedure or to more rapid catabolism of LTC4 and LTD4 by bronchial secretions in asthma than in cystic fibrosis.
对支气管哮喘、慢性支气管炎和囊性纤维化患者的痰液样本进行检查,以确定其中是否存在白三烯B4、C4和D4。在乙醇提取并通过Amberlite XAD - 8纯化后,使用合适的标志物通过高压液相色谱法(HPLC)鉴定白三烯。还使用博伊登趋化试验和豚鼠离体回肠FPL 55712可抑制的收缩对HPLC分离的组分进行生物活性测试。在支气管哮喘(7例中的7例)、慢性支气管炎(4例中的4例)和囊性纤维化(4例中的4例)患者经HPLC分离的痰液中检测到LTB4。相比之下,对豚鼠回肠进行生物测定未能在17份哮喘患者痰液中检测到LTC4或LTD4,尽管在5例支气管炎患者中的1例以及25例囊性纤维化患者中的16例中检测到了它们。对其中8份囊性纤维化痰液样本的活性通过HPLC进一步表征,结果显示为LTC4和/或LTD4。17例哮喘患者中的11例、25例囊性纤维化患者中的4例以及5例支气管炎患者中的2例的痰液中含有类过敏毒素物质。大多数含有LTB4的痰液样本还具有5(S),12(S),6 - 反式LTB4异构体的物理和生物学特性的活性。这些研究表明,花生四烯酸代谢的脂氧合酶产物以多种形式存在于各种阻塞性气道疾病的痰液中。在支气管哮喘患者痰液中未能检测到“慢反应物质 - A”白三烯,可能归因于提取过程中的损失、检测方法的不敏感性,或者是哮喘患者支气管分泌物对LTC4和LTD4的分解代谢比囊性纤维化患者更快。