抗TNF-α、IL-1βIgY抗体抗豚鼠过敏性支气管哮喘的实验研究

The experimental study on IgY against tumour necrosis factor alpha and interleukin-1beta allergic bronchial asthma of guinea pigs

目的:探讨抗TNF-α、IL-1βIgY抗体治疗过敏性支气管哮喘的作用机制。方法:卵清白蛋白雾化吸入制备Hart-ley豚鼠过敏性支气管哮喘动物模型,随机分正常对照组(A组)、过敏性支气管哮喘模型组(B组)、0.1%抗TNF-α、IL-1βIgY(C组)和1.0%抗TNF-α、IL-1βIgY雾化吸入治疗组(D组)。各组豚鼠治疗结束后2、4、8、24小时处死,肺组织切片H.E.染色;收集支气管肺泡灌洗液(BALF)Wright’s染色计数炎症细胞。结果:①肺组织病理改变:B组各时间点可见肺泡管和肺泡壁结构受损,肺泡腔内充满渗出液、上皮细胞和白细胞,肺间质水肿、炎性细胞浸润、毛细血管扭曲或扩张淤血、有效血流的毛细血管显著减少。C组和D组肺泡管和肺泡壁结构受损轻于B组,肺泡腔内偶见少量炎症细胞,支气管腔及肺泡中的粘液栓较B组明显减少,支气管周围少见炎性细胞浸润,支气管粘膜上皮修复现象明显。②BALF细胞学变化:与B组相比,C和D组嗜酸性粒细胞、中性粒细胞、淋巴细胞数量显著下降(2、4、8小时,P均<0.05);巨噬细胞显著上升(2、4、8小时,P均<0.05)。结论:抗TNF-α、IL-1βIgY抗体雾化吸入治疗豚鼠过敏性支气管哮喘能够显著减轻炎症病理反应。0.1%与1.0%的抗TNF-α、IL-1βIgY抗体疗效相当。

Objective：To investigate the mechanism of immunoglobulin Y antibodies（IgY） against tumour necrosis factor alpha（TNF-α） and Interleukin-lbeta（IL-1β） in treating allergic bronchial asthma through nebulization inhalation. Methods： The allergic bronchial asthma model was established with Hartley guinea pig by ovalbumin nebulizating inhalation. The animals were randomly divided into 4 groups： normal control group（ group A）, allergic bronchial asthma group （ group B）, 0.1% anti-TNF-α and IL- 1β IgY treating group （ group C）, 1.0 % anti- TNF-α and IL-1β IgY treating group （group D） .The animals were killed after treatment being accomplished for 2 h,4 h,8 h,24 h and the lungs were made pathological, which were then stained by hematoxylin-eosin（ H.E. ）. The bronchoalveolar lavage fluid （BALF） was collected and the deposited cells were stained by Wright＇ s. Results： ①The histological appearance of lung： In group B the histological structure of alveolar ducts and alveolar walls was damaged, the alveolar space was full of transudate and lots of alveolar epithelial cells and leucocytes. The pulmonary interstitial edema, inflammatory cells infiltration, distorted or dilated capillaries and reducing capillary numbers of effective blood stream were observed in alveolar walls. In group C and group D the damage degree of alveolar ducts and alveolar wall was slighter than that in group B and there were few inflammatory cells in alveolar space. In bronchial lumen and pulmonary alveoli the sticky mucus plug was obviously less in group C and group D than in group B. Moreover, inflammatory cell infiltration was seldom observed aroud brenchia, and restoration of bronchial tunica mucosa epithelium was obviously observed in group C and group D. ②The cytology appearance of BALF： In group C and group D the numbers of eosinophils, neutmphils, lymphocytes were significantly fewer （2 h, 4 h, 8 h, P 〈 0.05）, however, the number of macmphage was significantly more（2 h,4 h,8 h,P 〈 0.05）than in group B. Conclusion：The anti-TNF-α and IL-1β IgY can obviously alleviate pathological extent of inflammatory reaction in allergic bronchial asthma of guinea pigs by nebulization inhalation therapy. The therapeutic effect of anti-TNF-α and IL- 1β IgY between 0. 1% and 1.0% concentration is not obvious difference for pathology changes.