蛋白芯片技术分析 BALB/c小鼠 AD模型中皮损细胞因子表达的实验研究

摘要

Objective To investigate the expression of cytokine in atopic dermatitis model of BALB/c mouse which induced by topical application of DNCB and OX. Methods The induction of AD-like skin lesion was per￣formed by topical application of DNCB and OX. The clinical features,scratching frequency,the ear thickness and to￣tal serum IgE was measured to assess the AD-like condition,after different time point later of the first induction. Pro￣tein chip technology was used to detect the level of various cytokines,which include IL-4, IL-5, IL-13, IL-12,IFN-r,IL-2,IL-17,IL-10,TNF-αand ICAM-1.Results The results presented here have shown that 1%DNCB and 1%OX was repeatedly applied on the ear of BALB/c mice to induce AD-like lesion, including erythema, ede￣ma,scaling and excoriation with increased scratching behavior. Significant ear swelling was observed and the total se￣rum IgE was increased. AD-like histological features also was observed,which characterized in increased epidermal thickness,telangiectasis and infiltration of inflammatory cells in super-dermis. Up-regulated expression of IL-4,IL-5,IL-13,IL-17,IL-10,TNF-αand ICAM-12, and down-regulated expression of IL-12, IFN-r and IL-2was also observed in AD-like lesion.Conclusion It suggested that repeated application of DNCB and OX was efficient for induction of BALB/c mice AD model.%目的研究1％DNCB与1％OX经皮致敏诱发BALB/c小鼠双耳皮损的特点及小鼠炎症因子定量抗体芯片方法测定皮损中细胞因子表达水平，为开发药物的需要建立一种简单而又可重复的AD动物模型。方法将36只BALB/c小鼠脱毛后，分正常组、丙酮组、模型组各12只。1％DNCB和1％OX经皮致敏诱发模型组BALB/c小鼠，于7d、14d、21d、28d各处死3只小鼠并观察致敏皮损、搔抓情况及鼠耳厚度变化，小鼠IgE ELISA Kit检测血清总IgE水平，HE染色组织病理改变，小鼠炎症因子定量抗体芯片检测皮损中炎症因子(IL－4、IL－5、IL－13、IL－12、IFN－r、IL－2、IL－17、IL－10、TNF－α、ICAM－1)的含量。结果7d、14d、21d、28d后正常组和丙酮组鼠耳无明显变化，而1％DNCB和1％OX经皮致敏BALB/c小鼠鼠耳开始出现红斑、鳞屑及血痂；模型组小鼠出现搔抓，与正常组及丙酮组比较，P均<0�05，差异具有统计学意义；鼠耳明显增厚，与正常组及丙酮组比较，P均<0�05，差异具有统计学意义；组织病理模型组表皮增厚，毛细血管扩张，可见中性粒细胞、淋巴细胞浸润；模型组皮损中炎症因子IL－4、IL－5、IL－13、IL－17、IL－10、TNF－α、ICAM－12含量增多，IL－12、IFN－r、IL－2皮损中含量减少，其中IL－4、1L－5、IL－13、IL－2、IL－10及ICAM－1与正常组及丙酮组比较差异具有统计学意义( P均<0�05)，而IL－12、IFN－r、IL－17、TNF－α比较，差异无统计学意义( P均>0�05)。结论1％DNCB和1％OX经皮致敏可诱发BALB/c小鼠Th2优势型AD模型，BALA/c模型鼠皮损中IL－4、IL－5、IL－13含量水平增高，同时IL－17、IL－10、TNF－α、ICAM－12含量增高，IL－12、IFN－r、IL－2皮损中含量减少。