不同条件下建立过敏性结膜炎小鼠模型的比较研究  

Comparison of mouse models of allergic conjunctivitis under different conditions

作　　者：白梦天 李韵 胡竹林[1,2,3,4] BAI Mengtian;LI Yun;HU Zhulin(Department of Ophthalmology,Fourth Affiliated Hospital of Kunming Medical University,Kunming 650032,China;Yunnan Eye institute,Kunming 650032;Yunnan Second People’s Hospital,Kunming 650032;Expert workstation of Yao Ke,Kunming 650032;the First Affiliated Hospital of Kunming Medical University,Kunming 650032)

机构地区：[1]昆明医科大学第四附属医院眼科,昆明650032 [2]云南省眼科研究所,昆明650032 [3]云南省第二人民医院眼科,昆明650032 [4]姚克专家工作站,昆明650032 [5]昆明医科大学第一附属医院,昆明650032

出　　处：《中国实验动物学报》2020年第3期330-337,共8页Acta Laboratorium Animalis Scientia Sinica

基　　金：云南省眼科疾病防治研究重点实验室(2017DG008);云南省第二人民医院白内障与眼底疾病防治省创新团队(2017HC010);云南省姚克专家工作站(2017IC064)。

摘　　要：目的分别在不同抗原、不同激发途径、不同小鼠周龄条件下,建立过敏性结膜炎小鼠模型,通过比较免疫应答状态,筛选最佳致敏条件,为过敏性结膜炎小鼠动物模型的建立提供新的思路和方法。方法 SPF级Balb/c小鼠65只,在不同抗原(卵清蛋白、豚草花粉、屋尘螨),不同激发途径(灌胃、雾化吸入、皮下注射),不同小鼠周龄(1周龄、2周龄、3周龄、8周龄)条件下构建过敏性结膜炎小鼠动物模型。RT-qPCR检测小鼠结膜组织中CCL5 mRNA、IL-17 mRNA相对表达水平,流式细胞术检测脾脏单细胞悬液IL-17占CD4^+T细胞百分比。结果同阴性对照组比较,卵清蛋白组、屋尘螨组、豚草花粉组IL-17百分比、CCL5 mRNA及IL-17 mRNA表达均上升,其中,以豚草花粉组上升的最为显著。同阴性对照组比较,皮下注射组、灌胃组、雾化吸入组、阳性对照组IL-17百分比、IL-17 mRNA表达均上升,CCL5 mRNA在灌胃组中的表达与阴性对照组接近,皮下注射组的表达水平更接近阳性对照组。1周龄、2周龄组小鼠IL-17百分比、CCL5 mRNA、IL-17 mRNA表达水平较为接近,但显著低于3周龄、8周龄组小鼠。结论豚草花粉在构建过敏性结膜炎小鼠动物模型中具有更好的致敏性。皮下注射法激发抗原在过敏性结膜炎小鼠动物模型构造中可取得较满意效果,尽管未见明显眼部症状及体征,但全身和局部均呈致敏状态。3周龄Balb/c小鼠免疫应答反应已较为接近成年Balb/c小鼠。Objective To establish a mouse model of allergic conjunctivitis by comparison of different antigens,routes of administration,and ages of mice.We compared their immune response status and screened for optimal sensitization conditions to provide new ideas and method to establish mouse models of allergic conjunctivitis.Methods Sixty-five SPF Balb/c mice were selected to establish allergic conjunctivitis models using different antigens(ovalbumin,ragweed pollen,house dust mite),administration routes(gastric administration,aerosol inhalation,subcutaneous injection)and ages(1,2,3 and 8 weeks).Results The expression levels of interleukin-17(IL-17)and C-C motif chemokine ligand 5(CCL5)mRNA were increased in the ovalbumin,house dust mite and ragweed pollen groups,especially the later,compared with the negative control group.Compared with the negative control group,the expression of IL-17 mRNA was increased in the subcutaneous injection,gavage,inhalation and positive control groups.The expression of CCL5 mRNA in the gavage group was close to that of the negative control group,while its expression level in the subcutaneous injection group was closer to that of the positive control group.Expression levels of CCL5 and IL-17 mRNA were similar in 1-and 2-week-old mice,but significantly lower than those in 3-and 8-week-old mice.Conclusions Ragweed pollen provided better sensitization in the preparation of an allergic conjunctivitis mouse model.Subcutaneous injection of antigen achieved satisfactory result in the allergic conjunctivitis mouse model.Although there were no obvious eye symptoms,the whole body and local areas were sensitized.The immune response of 3-week-old Balb/c mice was close to that of adult Balb/c mice.

关 键 词：过敏性结膜炎 白介素17 屋尘螨 卵清蛋白 豚草花粉 

分 类 号：Q95-33[生物学—动物学]