吸入水雾剂的过敏性试验方法研究

目的：建立与吸入水雾剂临床给药方式相似，可用于评价吸入水雾剂过敏性的实验方法。方法：Hartley豚鼠24只，雌雄各半，随机分为阴性对照组及3个给药组。给药组于1、3、5 d按每只2 mL吸入浓度为1%的雾化卵白蛋白（ovalbumin，OVA）致敏，阴性对照组以氯化钠注射液（NS）同法处理。于首次致敏后14 d将给药组分别以0.5%、1%或2%OVA激发，阴性对照组以NS代替OVA同法处理。激发后观察豚鼠出现的过敏症状，取血检测血液中嗜酸性粒细胞（eosnophils，EOS）比例。取肺泡灌洗液（bronchoalveolar lavage fluid，BALF）检测组胺（histamine，HIS）水平。解剖豚鼠，大体观察后取鼻粘膜、气管和肺进行病理学检查。结果：给药组豚鼠出现不同程度过敏症状及相应病理学改变，过敏反应严重程度、EOS比例及HIS水平均与激发液OVA浓度相关。结论：豚鼠吸入致敏与激发的过敏性实验方法能反映典型过敏症状，方法稳定、简易，能较好地应用于吸入水雾剂的过敏性评价。

Objective: To establish an allergy test method for evaluating the allergy of aqueous aerosol inhalation. Methods: A total of 24 hartley guinea pigs were randomly divided into negative control group and three medication groups in which normal saline and 1% ovalbumin (OVA) were administrated to related animals by inhalation on Day 1,Day 3 and Day 5 for sensitization.On Day 14,animals in medication groups were challenged with 0.5%,1% or 2% OVA by inhalation,and negative control group with normal saline.Then the occurrence of allergic symptoms was recorded,the blood samples were collected to examine the percentage of eosnophils (EOS) in blood,and the histamine level in bronchoalveolar lavage fluid (BALF) was analyzed,and the pathological examination of nasal mucosa,trachea and lung was performed after macroscopic observation on dissected guinea pigs. Results: Animals in medication groups showed typical allergic symptoms and related pathologic changes.The severity of allergic symptoms and the levels of EOS and HIS in medication groups were all associated with the OVA concentration. Conclusion: The method sensitizing and challenging guinea pigs by inhalation is easy and stable,which reflects the typical allergy symptoms.It is suitable for evaluating the allergy of aqueous aerosol inhalation.

[1] CHEN Gui-liang(陈桂良), HOU Hui-min(侯惠民).The effectiveness and quality evaluation of the dry power inhaler(干粉吸入剂的有效性及质量评价)[J].Chin J Pharm(中国医药工业杂志), 2004, 35(1):50
[2] Weda M, Zanen P, de Boer AH, et al.Equivalence testing of salbutamol dry powder inhalers:in vitro impaction results versus in vivo efficacy[J].Int J Pharm, 2002, 249(1-2):247
[3] Ashurst IC, Herman CS, Li-Bovet L, et al.Metered Dose Inhaler for Albuterol:US, 6131566[P].2000-10-17
[4] Nabe T, Mizutani N, Osaki S, et al.Comparison of ceflal-polleninduced allergic rhinitis in passively and actively sensitized guinea pigs[J].Jpn J Pharmacol, 2001, 85(4):409
[5] DI Chan-juan(邸婵娟), LI Hai-bing(李海兵), HAN Chun-guang(韩春光).Characteristics and identification of an ovalbumin-induced aller gic rhinitis model in guinea pigs(卵白蛋白诱发变应性鼻炎豚鼠模型的特点及鉴定)[J].Carcinog Teratog Mutag (癌变·畸变·突变), 2012, 24(6):440
[6] YAN Cui-xia(严翠霞), JIANG Wen-ming(江文明), CHEN Gui-liang(陈桂良), et al.Critical evaluation of determination methods of the particle size of dry powder inhaler:a comparison of three impactors(吸入粉雾剂粒径测定方法的评价:3种撞击器的比较)[J].Chin J Pharm Anal(药物分析杂志), 2011, 31(7):1296
[7] WANG Chang-zheng(王长征).Eosinophils and bronchial asthma(嗜酸性粒细胞与支气管哮喘)[J].Chin J Tuberc Respir Dis(中华结核和呼吸杂志), 1998, 21(9):562
[8] Peng T, Hao L, Madri JA, et al. Role of C5 in the development of airway inflammation, airway hyperresponsiveness, and ongoing airway response[J].J Clin Invest, 2005, 115(6):1590
[9] ZHANG Fang-qi(张方琪), YANG Xue-min(杨学敏), TANG Yuan-yuan(唐元元).Research progress of eosinophils in the pathogenesis of asthma(嗜酸性粒细胞在哮喘发病机制中的研究进展)[J].Chin J Lung Dis(中华肺部疾病杂志), 2013, 6(2):162
[10] Kay AB.Asthma and inflammation[J].J Allergy Clin Immunol, 1991, 87(5):893
[11] Takeuchi K, Kishioka C, Ishinaga H, et al.Histamine alters gene expression in cultured human nasal epithelial cells[J].J Allergy Clin Immunol, 2001, 107(2):310
[12] Kohda F, Koga T, Uchi H, et al.Histamine-induced IL-6 and IL-8 production and differentially modulated by IFN-gamma and IL-4 in human keratinocytes[J].J Dermatol Sci, 2002, 28(1):34
[13] Yamasaki ML, Sasaki K, Mizutani N, et al.Pharmacological characterization of the leukocyte kinetic after intranasal antigen challenge in a guinea pig model of allergic rhinitis[J].Inflamm Res, 2001, 50(9):474