白介素17单克隆抗体对变应性鼻炎小鼠气道炎症的作用

Effect of Interleukin-17 Monoclonal Antibody on Airway Inflammation in Mice with Allergic Rhinitis

目的探讨白介素17单克隆抗体(IL-17mAb)的不同给予剂量及方式在变应性鼻炎小鼠气道炎症中的作用。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 d将20μg卵清蛋白(OVA)加2 mg铝佐剂腹腔注射处理A、C、D、E及F组小鼠,间隔7 d,第22天开始进行鼻腔激发,每天每侧鼻孔各给予OVA 10μl(共500μg)滴鼻,连续7 d。A、C、D、E组小鼠于每次OVA鼻腔激发前1 h分别给予生理盐水、100 ng IL-17mAb、500 ng IL-17mAb、5μg IL-17mAb滴鼻,F组小鼠于每次OVA鼻腔激发前4 h给予5μg IL-17mAb腹腔注射,B组小鼠于相同时间点给予等量生理盐水腹腔注射及滴鼻。所有小鼠于最后1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方法检测血清及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。结果4周末A组所有小鼠症状学评分均>5分,提示造模成功。F组小鼠的挠鼻及喷嚏次数均少于A组(P<0.05);F组小鼠NLF中嗜酸性粒细胞数、血清IL-6水平低于A组,血清及NLF中IL-10水平均高于A组(P<0.05);E组小鼠血清中IL-10水平高于A组(P<0.05);A组小鼠鼻黏膜组织中肥大细胞数多于B组,统计学意义显著(P<0.01);F组小鼠鼻黏膜组织中肥大细胞数少于A组,但差异无统计学意义(P>0.05);F组小鼠鼻黏膜组织中肥大细胞数与B组比较,差异无统计学意义(P>0.05)。结论高剂量的(5μg)IL-17mAb腹腔注射处于激发阶段的变应性鼻炎小鼠促使小鼠变应性鼻炎症状明显减轻,鼻腔灌洗液嗜酸性粒细胞减少。促使变应性鼻炎小鼠血清中IL-6表达降低,血清中及鼻腔灌洗液中IL-10表达升高,因此推测这些细胞因子的变化可能抑制Th17/促进Treg的分化,进而对变态反应产生抑制作用。

Objective To investigate the effects of different doses and modes of interleukin-17 monoclonal antibody(IL-17mAb)on airway inflammation in mice with allergic rhinitis.Methods 48 mice were randomly divided into groups A,B,C,D,E,and F,with 8 rats in each group.On the 0th,7th,and 14th d,20μg of ovalbumin(OVA)plus 2 mg of aluminum adjuvant were intraperitoneally injected into mice of group A,C,D,E and F at intervals of 7 d.The nasal challenge began on the 22nd d.10μl(500μg total)of OVA was administered to each nostril on each day for 7 d.Group A,C,D,and E mice were given normal saline,100 ng IL-17 mAb,500 ng IL-17 mAb,and 5μg IL-17 mAb nasally 1 h before each OVA nasal challenge.Group F mice were given.5μg of IL-17mAb was intraperitoneally injected 4 h before OVA nasal challenge,and group B mice were given the same amount of normal saline intraperitoneal injection and nasal drops at the same time point.All mice were evaluated for nasal symptom changes after the last challenge.Diff-Quik staining was used to observe eosinophil infiltration in nasal lavage fluid(NLF).ELISA was used to detect the level of IL-6 and IL-10 in serum and NLF.The nasal mucosa was stained with toluidine blue for observation of mast cells.Results After 4 weeks,all mice in group A had a symptoms score of>5,suggesting successful modeling.The number of sneezing and sneezing in group F was lower than that in group A(P<0.05).The number of eosinophils in NLF and serum IL-6 of group F were lower than those in group A,IL-10 in serum and NLF of group F was higher than that of group A(P<0.05).The level of IL-10 in serum of group E was higher than that of group A(P<0.05).The number of mast cells in nasal mucosa of group A was higher than that of group B,the difference was significant(P<0.01).The number of mast cells in nasal mucosa of group F was less than that of group A,but the difference was not statistically significant(P>0.05).The number of mast cells in nasal mucosa of group F was similar to group B,there was no significant difference in the B group(P>0.05).Conclusion High-dose(5μg)IL-17mAb intraperitoneal injection of allergic rhinitis mice in the stimulating phase promotes a significant reduction in allergic rhinitis in mice and a decrease in eosinophils in nasal lavage fluid.Decrease the expression of IL-6 in the serum of mice with allergic rhinitis,and increase the expression of IL-10 in serum and nasal lavage fluid.Therefore,it is speculated that changes in these cytokines may inhibit Th17/promoting the differentiation of Treg,thereby inhibiting allergic reactions.