镍变应性接触性皮炎豚鼠动物模型的构建

摘要

Objective To establish a guinea-pig model of nickel allergic contact dermatitis (ACD ) and investigate the influence of different experimental conditions (such as the concentration of nickel,duration of exposure,cutaneous microenvironment,and the schedule of elicitation)on the establishment the nickel ACD model.Methods Eighty-six guinea-pigs (240-270 g)were divided randomly into fourteen groups based on different factors (the concentration of nickel,duration of exposure,cutaneous microenvironment, and the schedule of elicitation).The 20%Ni-28d-6h,1 0%Ni-28d-6h,20%Ni-28d-SP-6h,1 0%Ni-28d-SP-6h,20% Ni-28d-24h,1 0% Ni-28d-24h,20% Ni-28d-SP-24h,and 1 0% Ni-28d-SP-24h group was sensitized in day 0,7,1 4 and challenged in day 28 with nickel sulfate,whereas,the 20%Ni-9d-6h,20%Ni-9d-24h,20%Ni-9d-SP-6h,20% Ni-9d-SP-24h,positive control (PC),and negative control (NC) group was sensitized in day 0,1 ,2 and challenged in day 9.Dynamic grading of the skin alterations was performed at 6,1 2,24,48,and 72 h after challenge.The levels of IL-2 and IFN-γin the guinea pig’s skin of group 20%Ni-9d-6h,20%Ni-9d-24h,20%Ni-9d-SP-6h,20%Ni-9d-SP-24h,PC,and NC was detected by ELISA methods.H.E staining was used for the pathological biopsy.Results The most severe erythemaic reaction was observed at 48 h in the groups of day 28,whereas at 24 h in the groups of day 9. Swelling was not obvious in all the guinea pigs.The levels of IL-2 and IFN-γin the skin lesions increased to the peak level (IL-2 533.53 ±41 .51 pg/ml),(IFN-γ1 235.63 ±76.85 pg/ml)at 48 h in the groups of day 9 untill 72 h.Inflammatory cells infiltrating,escharosis,hyperkeratosis,angiotelectasis was shown in the epidermis and dermis in all the animals,which is similar to human ACD.Conclusions The guinea-pig models of nickel ACD had been successfully established for different experimental conditions.The higher nickel level and resulted in more severe duration of exposure longer skin reaction.Similar results can be achieved based on synthetic perspiration (SP)in the nickel and the schedule of sensitizing in day 0,1 ,2 and challenged in day 9.%目的：建立豚鼠镍变应性接触性皮炎（allergic contact dermatitis，ACD）的动物模型，观察不同条件（镍浓度、致敏时间、汗液和接触时间）对造模的影响，为临床前评价镍 ACD 治疗药物提供动物模型。方法选择86只240～270 g 豚鼠，按不同条件（硫酸镍浓度、致敏时间、人工汗液、封闭接触时间）随机分为14组，第0天、第7天、第14天致敏且第28天激发的8组（20％Ni-28d-6h、10％Ni-28d-6h、20％Ni-28d-SP-6h、10％Ni-28d-SP-6h、20％Ni-28d-24h、10％Ni-28d-24h、20％Ni-28d-SP-24h、10％Ni-28d-SP-24h），第0天、第1天、第2天连续3天致敏且第9天激发的6组（20％Ni-9d-6h、20％Ni-9d-24h、20％Ni-9d-SP-6h、20％Ni-9d-SP-24h、阳性对照组、阴性对照组）；激发后0～72 h，对激发部位皮肤红斑和水肿情况进行动态分级；分别于激发后24、48、72 h 采集第9天激发的6组豚鼠激发部位皮肤，酶联免疫吸附法检测皮损组织中白介素（interleukin，IL）-2和干扰素（interferon，IFN）-γ含量；HE 染色观察激发部位皮肤病理变化；结果第28天激发组，激发后各组激发部位皮肤48 h 红斑程度最强，并于48 h 后红斑程度减轻；第9天激发组，红斑在24 h 程度最强并一直持续到72 h；而皮肤水肿各组均不明显。第9天激发后皮损部位皮肤组织中 IL-2和 IFN-γ均在48 h 达到峰值并持续到72 h，峰值分别为（533．53±41．51）pg/ml 和（1235．63±76．85）pg/ml。激发部位皮肤表面结痂，表皮增厚，轻度角化过度，真皮毛细血管扩张，真皮及表皮内有炎性细胞浸润，与人类 ACD 病理表现一致。结论第0天、第1天、第2天致敏且第9天激发的致敏方法能建立皮肤反应较严重的 ACD 模型，并通过增加镍浓度、封闭接触时间以及人工汗液控制变态反应程度；第0天、第7天、第14天致敏且第28天激发的致敏方法则能得到不同程度的皮肤反应 ACD 模型；本研究为研究镍 ACD 提供了与临床相似的各种豚鼠 ACD 动物模型。