研究天山雪莲多糖对特异性皮炎(AD)小鼠的作用及调控机制。外用2,4-二硝基氟苯(DNFB)反复诱导建立昆明小鼠AD模型,40只小鼠,每组8只,随机分为:空白组、模型组、阳性泼尼松龙组和天山雪莲多糖剂量组(质量浓度为100和500 μg/mL)。记录各组小鼠皮损情况,检测小鼠经皮失水率,观察皮损组织的病理学,ELISA法测定天山雪莲多糖高剂量组皮损组织中IL-6,IL-4,IL-13,Ig E,TNF-a,NOD 1,TLR-2,CRAMPS的水平。相对于模型组,外用天山雪莲多糖处理小鼠皮肤特异性皮炎湿疹样得到明显改善,经皮失水率显著降低,HE染色病理图片的表皮层厚度显著降低,皮肤组织中Th2相关的细胞因子IL-6,IL-4,IL-13和Ig E的表达均显著下降(P<0.01),同时TLR受体调控相关的因子NOD 1,TLR-2,TNF-α的蛋白表达均显著降低(P<0.01),CRAMPS含量显著升高(P<0.05)。结果表明,天山雪莲多糖通过调控TLR-2和Th2相关细胞因子的表达,改善了AD模型小鼠的皮损,为天山雪莲多糖在预防和治疗特异性皮炎制剂中的应用提供数据支持。
The therapeutic effect and its possible mechanism of Saussurea involucrata polysaccharide on atopic dermatitis (AD) in mice were studied. The polysaccharide of Saussurea involucrata was obtained by water extraction and alcohol precipitation process. The content of crude polysaccharide was determined. The atopic dermatitis model in mice was established with topical 2, 4-dinitrofluorobenzene (DNFB). 40 Mice were randomly divided into blank group, model group, prednisolone positive group, and low-dose and high-dose Saussurea involucrata polysaccharide groups (100 and 500 μg/mL, respectively). The skin lesions of each group of mice were recorded; the transepidermal water loss of the mice was tested; the lesions were stained with hematoxylin-eosin to observe the pathological characteristics; the expression levels of IL-6, IL-4, IL-13, Ig E, TNF-α, NOD 1, TLR-2 and CRAMPS in the skin lesions were determined by ELISA method. The results show that the content of Saussurea involucrata polysaccharide prepared in this experiment is 28.23%. Saussurea involucrate polysaccharide could effectively alleviate the skin symptoms of mice with DNFB-induced atopic dermatitis. The transepidermal water loss and the thickness of the epidermal layer are both reduced. The high-dose group of Saussurea involucrata polysaccharide was selected for cytokine detection. Compared with the model group, the expression levels of Th2-related cytokines IL-6 (P<0.01), IL-4 (P<0.01), IL-13 (P<0.01) and Ig E in the skin of mice (P<0.01) are effectively reduced for the Saussurea involucrate polysaccharide group. Meanwhile, the Saussurea involucrate polysaccharide group regulates the cytokines of the innate immune pathway, significantly reduces the expression levels of TLR-2 (P<0.01), NOD1 (P<0.01) and TNF-α (P<0.01), and significantly increases the expression level of CRAMPS (P<0.05). The present study shows that, through inhibiting the cytokines IL-4 and IL-13 in Th2 inflammatory response process in atopic dermatitis, and through regulating the secretion and release of the innate immunity regulator of TLR-2, the inflammatory response in AD model mice is thereby inhibited.