变应性鼻炎动物模型建立及白细胞介素6、10因子检测的初步报告

Establishment of allergic rhinitis animal model and its application in detection of interleukin 6 and 10

目的建立变应性鼻炎(allergic rhinitis,AR)豚鼠模型并检测白细胞介素6(interleukin 6,IL-6)和白细胞介素10(interleukin 10,IL-10)。方法选用20只Hartley豚鼠,实验组和对照组各10只,随机分配。卵清蛋白(ovalbumin,OVA)作为致敏原,氢氧化铝作为佐剂。分两个阶段进行实验,第一阶段全身致敏,第二阶段鼻部激发。实验组OVA混悬液1ml腹腔注射,隔日1次,共7次;对照组0.9%氯化钠注射液1 ml腹腔注射。全身致敏后隔3 d用OVA溶液滴鼻,每侧鼻腔0.05 ml,1次/d,连续7 d;对照组用0.9%氯化钠注射液滴鼻。观察鼻痒(挠鼻)、喷嚏、清涕、哮喘等行为,用1~3分对四项行为评分,总分5分以上判定造模成功。高倍病理显微镜下计数嗜酸性粒细胞,用试剂盒检测黏膜组织中IL-6和IL-10。结果实验组豚鼠被致敏和激发后,频繁出现鼻痒(挠鼻)、喷嚏、清涕等行为,实验组造模成功率60%,实验组较对照组鼻黏膜嗜酸性粒细胞浸润明显增多(P<0.05)。实验组死亡1只,其肺组织以及支气管黏膜较对照组嗜酸性粒细胞浸润明显。实验组豚鼠鼻黏膜组织IL-6水平为(185.41±9.57)ng/L,IL-10水平为(99.97±12.71)ng/L,较对照组IL-6水平(4.35±0.30)ng/L、IL-10水平(4.45±0.31)ng/L明显增高(P均<0.05)。结论 AR豚鼠模型成功建立,为后续研究奠定了基础。

Objective To establish guinea pig model for allergic Rhinitis（AR） and detect interleukin 6（IL-6） and interleukin 10（IL-10） in it. Methods Twenty Hartley guinea pigs were selected and randomly divided into experimental group and control group with each group of 10 animals. Ovalbumin（OVA） was used as allergen and aluminum hydroxide as adjuvant. The experiment included two stages,which were systemic sensitization and nose provocation,the experimental group was induced by intraperitoneal injection with OVA suspension 1 ml,once every two days for 7 times,and the control group was induced by intraperitoneal injection with normal saline 1 ml once every two days for 7 times. Three days after systemic sensitization,OVA solution was dropped into nose with 0.05 ml for each nasal cavity,once a day for 7 days. The behaviors such as nasal itching,sneezing,nasal discharge,asthma were carefully investigated. The behaviors were scaled from 1 to 3 scores and more than 5 scores in total were considered as a successful model. The eosinophils was counted under high magnification microscope（×400）,and IL-6 and IL-10 were detected by ELISA Kits. Results The guinea pigs in experimental group after sensitizing and provocation developed frequent nasal itching,sneezing and rhinorrhea. The success rate of animal model in the experimental group was 60%. Compared with control group,eosinophil infiltration in experimental group increased significantly（P〈0.05）. The dead guinea pig in experimental group showed significant eosinophil infiltration in lung and bronchial mucosa compared to the control group seemingly like asthmatic inflammation. The experimental group had higher IL-6 level（185.41±9.57） ng/L,IL-10 level（99.97±12.71） ng/L than those in control group [IL-6 level（4.35±0.30） ng/L,IL-10（4.45±0.31） ng/L]（P〈0.05,respectively）. Conclusion The guinea pig model for AR is successfully established and lays the foundation for the following studies.