卵清蛋白、金黄色葡萄球菌肠毒素B和维生素D类似物诱导小鼠特应性皮炎模型

摘要

目的:阐明利用卵清蛋白(ovalbumin,OVA)、金黄色葡萄球菌肠毒素B(staphylococcal enterotoxin B,SEB)和维生素D类似物(calcipotriene ointment,CO)局部刺激小鼠背部建立特应性皮炎(atopic dermatitis,AD)动物模型的方法.方法:将BALB/c小鼠背部去毛随机分为3组:对照组,OVA/SEB,OVA/SEB/CO,分别给予相应的处理.隔天给药并拍照记录皮损状态,于第15天处死小鼠,处死前用皮肤共聚焦显微镜测量表皮厚度.用ELISA方法检测血清IL-2,IL-4,IL-31,干扰素(interferon,IFN)-γ,肿瘤坏死因子(tumor necrosis factor,TNF)-α,神经生长因子(nerve growthfactor,NGF)和IgE水平,HE染色检测皮损处炎症情况,实时荧光定量PCR检测皮损处上述因子的mRNA表达水平.结果:OVA/SEB/CO组皮损处炎症细胞浸润和表皮增厚明显,血清IgE水平显著增高,血清和皮损处IL-4,IL-31,IFN-γ,TNF-α和NGF表达水平均显著高于对照组或OVA/SEB组.结论:OVA/SEB/CO局部刺激可短时间、低成本地诱导BALB/c小鼠皮肤AD样改变.%Objective:To illuminate a method for establishment of a cost-efficient atopic dermatitis (AD) mouse model by topical application of ovalbumin (OVA),super-antigen staphylococcal enterotoxin B (SEB),and calcipotriene ointment (CO) on the back of BALB/c mice.Methods:Experimental mice were topically treated with OVA/SEB or OVA/SEB/CO every other day during 15 days of induction.Clinical alterations on the skin area were monitored every other day.Epidermal thickness were measured by reflectance confocal microscope (RCM) before harvest.Inflammatory cells in skin biopsies were marked by hematoxylin-eosin (HE) staining.Blood sample and skin biopsies were measured by ELISA and quantitative real-time PCR to detect the expression of IL-2,IL-4,IL-31,interferon (IFN)-γ,tumor necrosis factor (TNF)-α pruritus-associated nerve growth factor (NGF),and serum IgE.Results:Human AD-like cutaneous local inflammatory reaction was characterized by the accumulation of inflammatory cells,increased epidermal thickness and serum IgE levels as well as Th1 cell-associated cytokines (IFN-γ,TNF-α),Th2 cell-associated cytokines (IL-4,IL-31),and NGF in the OVA/SEB/CO group compared with that in the normal control group or the OVA/ SEB group.Conclusion:OVA/SEB/CO can induce an AD-like mouse model with lower economic and time consumption.