“血虚风燥型”皮炎-湿疹样病变豚鼠模型的评价指标研究

Evaluation Index of Guinea Pig Model of "Xuexu Fengzao" Dermatitis-Eczema-like Lesions

目的探讨“血虚风燥型”皮炎-湿疹样病变豚鼠模型的客观指标,为科学评价该模型提供依据。方法建立“血虚风燥型”皮炎-湿疹样病变豚鼠模型,每周应用MPA6/10多功能皮肤测试仪测量皮肤角质层含水量(WCSC)和经皮肤水分流失量(TEWL),HE染色后观察豚鼠皮损组织病理改变,酶联免疫吸附测定(ELISA)法检测血清免疫球蛋白E(IgE)、白细胞介素-4(IL-4)、IL-17、CD4^(+)、CD8^(+)浓度,蛋白质免疫印迹(WB)法检测皮损组织水通道蛋白3(AQP3)表达情况,逆转录聚合酶链反应(RT-PCR)检测皮损组织AQP3mRNA表达情况。结果与正常组比较,皮炎-湿疹样病变豚鼠模型组豚鼠背部致敏区出现红斑、丘疹、苔藓样变和明显搔抓,皮肤WCSC减少(P<0.01)、TEWL增多(P<0.01),病理显示皮肤表皮角化过度,颗粒层、棘细胞层、基底层细胞和真皮层纤维结缔组织增生,炎性细胞浸润,血清IgE、IL-17、CD4^(+)浓度和CD4^(+)/CD8^(+)比值显著降低(P<0.01),IL-4、CD8^(+)浓度显著升高(P<0.01),AQP3mRNA及蛋白表达明显降低(P<0.01)。结论WCSC和TEWL,组织病理变化,AQP3mRNA及蛋白表达能够客观反应出“血虚风燥证”的“皮肤燥象”特征,血清IgE、IL-4、IL-17、CD4^(+)、CD8^(+)浓度反应出“皮炎-湿疹”部分病理机制,2组结合可以作为“血虚风燥型”皮炎-湿疹样病变豚鼠模型的客观评价指标。

Objective In order to explore the objective indicators of the dermatitis-eczema-like lesions guinea pig model of"Xuexu Fengzao"to provide basis for scientific evaluation of the model.Methods Guinea pig model of"Xuexu Fengzao"dermatitis-eczema-like lesions was established,to measure the water content of the stratum corneum(WCSC)and the amount of transepidermal water loss(TEWL)every week.After HE staining,the pathological changes of guinea pigs skin lesions were observed.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum immunoglobulin E(IgE),interleukin-4(IL-4),IL-17,CD4^(+)and CD8^(+)concentrations.Western blotting(WB)was used to detect aquaporin-3(AQP3)protein expression in skin lesions.Fluorescence quantitative polymerase chain reaction(PCR)detection of AQP3mRNA expression in skin lesions.Results Compared with the normal control group,the dermatitis-eczema-like lesion guinea pig model group showed erythema,papules,lichenoid changes and obvious scratching on the sensitized area on the back of the guinea pigs,and the WCSC decreased(P<0.01)and TEWL increased(P<0.01).Pathology showed hyperkeratosis of the skin epidermis,hyperplasia of granular layer,spinous cell layer,basal layer cells and dermal fibrous connective tissue,and inflammatory cell infiltration.AQP3mRNA and protein expression was significantly reduced(P<0.01).Conclusion WCSC and TEWL,histopathological changes,AQP3mRNA and protein expression can objectively reflect the"skin dryness"characteristics of"Xuexu Fengzao".Serum IgE,IL-4,IL-17,CD4^(+)and CD8^(+)reflects part of the pathological mechanism of"dermatitis-eczema".