不同剂量外源性白介素17A干预对变应性鼻炎小鼠气道炎症的影响

doi:10.6040/j.issn.1673-3770.0.2018.365

摘要/Abstract

摘要： 目的探讨不同剂量的外源性白细胞介素17A(IL-17A)干预对变应性鼻炎小鼠气道炎症的影响。方法将48只小鼠采用随机数字表法分为A、B、C、D、E、F组,每组8只。分别于第0、7、14天将溶于50 μL生理盐水的20 μg卵清蛋白(OVA)加2 mg铝佐剂腹腔注射处理D、E及F组小鼠,间隔7 d,于第22天开始鼻腔激发,每天每侧鼻孔各滴入OVA 10 μL,连续7 d。D、E及F组小鼠于每次OVA鼻腔激发前1 h分别给予生理盐水、IL-17A蛋白100 ng、IL-17A蛋白500 ng滴鼻,A、B及C组小鼠于相同时点予等量生理盐水腹腔注射并滴鼻,B、C组小鼠于每次生理盐水鼻腔激发前1 h分别给予IL-17A蛋白100 ng、IL-17A蛋白500 ng滴鼻。所有小鼠于最后一次激发后评估鼻部症状学变化,Diff-Quik染色观察鼻腔灌洗液(NLF)中嗜酸性粒细胞浸润情况,ELISA法检测血清及NLF中IL-6、IL-10水平,鼻黏膜组织行甲苯胺蓝染色观察肥大细胞数。结果 4周末D组所有小鼠症状学评分均>5分,造模成功。A、B、C、D、E及F组症状学评分分别为1.5(1.0~2.8)、2.0(1.0~3.0)、2.5(2.0~3.0)、7.0(6.3~8.0)、3.0(2.3~3.8)及8.0(7.0~8.8)分。E组小鼠挠鼻及喷嚏次数少于D组(H=21.375, P=0.033; H=20.250, P=0.049)。D组小鼠血清及NLF中IL-6水平高于A组(H=25.750, P=0.004; H=20.688, P=0.047),IL-10水平低于A组(H=22.875, P=0.016; H=20.625, P=0.048)。E组小鼠血清及NLF中IL-6水平与A组相比差异无统计学意义(H=19.875, P=0.068; H=8.125, P>0.999)。E组小鼠血清中IL-10水平高于D组(H=27.062, P=0.002)。F组小鼠血清及NLF中IL-6水平高于A组(H=22.250, P=0.008; H=28.688, P=0.001)。F组小鼠血清及NLF中IL-10水平与D组相比差异无统计学意义(H=13.062, P=0.930; H=0.500, P>0.999)。C组小鼠NLF中嗜酸性粒细胞数高于A组(H=20.688, P=0.047)。E组小鼠NLF中嗜酸性粒细胞数低于D组(H=21.188, P=0.037)。F组小鼠NLF中嗜酸性粒细胞数与D组相比差异无统计学意义(H=2.875, P>0.999)。D组小鼠鼻黏膜组织中肥大细胞数与A组相比增多(H=27.188, P=0.002)。E组小鼠鼻黏膜组织中肥大细胞数少于D组(H=20.938, P=0.042)。F组小鼠鼻黏膜组织中肥大细胞数与D相比差异无统计学意义(H=1.188, P>0.999)。结论 500 ng的外源性IL-17A能促使小鼠鼻腔嗜酸性粒细胞募集,100 ng的外源性IL-17A对变应性鼻炎小鼠模型气道炎症有保护作用。

关键词: 鼻炎,变应性, 白细胞介素17A, 气道炎症, 小鼠

Abstract: Objective To evaluate the effects of different doses of exogenous Interleukin-17A(IL-17A)on airway inflammatory responses in mice with allergic rhinitis. Methods Forty-eight BALB/c female mice were assigned randomly to six groups(A, B, C, D, E, and F), with eight mice per group. Groups D, E, and F received intraperitoneal OVA injection and nasal challenge, normal saline(NS), and different doses of IL-17A(100 ng and 500 ng)one hour before nasal challenge, respectively. Groups A, B, and C received intraperitoneal NS injection and nasal challenge, NS, and different doses of IL-17A(100 ng and 500 ng)one hour before nasal challenge, respectively. Nasal symptoms were assessed at 4 weeks. Serum, nasal lavage fluid(NLF), and nasal tissue were collected. Eosinophils in NLF were measured using Diff-Quik staining. IL-6 and IL-10 levels in serum and NLF were measured using enzyme-linked immunosorbent assay(ELISA). Mast cells in nasal mucosa were measured using toluidine blue staining. Results All mice in group D had a symptom score of greater than five points, demonstrating that the model was successful. Groups A, B, C, D, E, and F had average scores of 1.5(1.0-2.8), 2.0(1.0-3.0), 2.5(2.0-3.0), 7.0(6.3-8.0), 3.0(2.3-3.8), and 8.0(7.0-8.8), respectively. Rubbing times and number of sneezes were lower in group E than in group D(H=21.375, P=0.033, H=20.250, P=0.049). Levels of IL-6 in the serum and NLF were higher in group D than in group A(H=25.750, P=0.004; H=20.688, P=0.047). IL-10 levels in the serum and NLF were lower in group D than in group A(H=22.875, P=0.016; H=20.625, P=0.048). There were no significant differences in IL-6 levels in the serum and NLF between group E and group A(H=19.875, P=0.068; H=8.125, P>0.999). IL-10 levels in the serum were higher in group E than in group D(H=27.062, P=0.002). IL-6 levels in the serum and NLF were higher in group F than in group A(H=22.250, P=0.008, H=28.688, P=0.001). There were no significant differences in IL-10 levels in the serum and NLF between group F and group D(H=13.062, P=0.930; H=0.500, P>0.999). Eosinophils in the NLF were more abundant in group C than in group A(H=20.688, P=0.047). The number of eosinophils in group E was significantly lower than that in group D(H=21.188, P=0.037). There was no significant difference in the number of eosinophils in group F compared with that in group D(H=2.875, P>0.999). Mast cells in the nasal mucosa were more abundant in group D than in group A(H=27.188, P=0.002). The number of mast cells in the nasal mucosa was lower in group E than in group D(H=20.938, P=0.042). There was no significant difference in the number of mast cells between group F and group D(H=1.188, P>0.999). Conclusion 500 ng of exogenous IL-17A led to a modest proinflammatory effect in normal mice. Treatment with 100 ng exogenous IL-17A in mice with allergic rhinitis can protect against allergic airway inflammation.

Key words: Rhinitis, allergic, IL-17A, Airway inflammation, Mouse

中图分类号:

R765.2

姜晓丹,董庆喆,党志红,李慎玲,苗玉,赵涵,张念凯. 不同剂量外源性白介素17A干预对变应性鼻炎小鼠气道炎症的影响[J]. 山东大学耳鼻喉眼学报, 2018, 32(6): 73-78.

JIANG Xiaodan, DONG Qingzhe, DANG Zhihong, LI Shenling, MIAO Yu, ZHAO Han, ZHANG Niankai. Dose-dependent effects of exogenous IL-17A on airway inflammatory responses in mice with allergic rhinitis[J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2018, 32(6): 73-78.

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