紫草素对特应性皮炎小鼠TSLP/OX40L通路及Th1/Th2平衡的影响  

Effects of shikonin on TSLP/OX40L pathway and Th1/Th2 cell balance in atopic dermatitis mice

作　　者：吴金环[1] 郑宝勇[1] 张理涛[2] WU Jin-huan;ZHENG Bao-yong;ZHANG Li-tao(Department of Dermatology,Baodi Clinical College of Tianjin Medical University,Tianjin 301800,China;Department of Dermatology,the Affiliated Hospital of Tianjin Academy of Traditional Chinese Medicine)

机构地区：[1]天津医科大学宝坻临床学院皮肤科,301800 [2]天津市中医药研究院附属医院皮肤科

出　　处：《天津医药》2021年第9期949-954,共6页Tianjin Medical Journal

摘　　要：目的探究紫草素对特应性皮炎(AD)小鼠胸腺基质淋巴细胞生成素(TSLP)/OX40配体(OX40L)通路及辅助性T细胞(Th)1/Th2平衡的影响。方法采用局部外涂2,4-二硝基氟苯(DNFB)法制备AD小鼠模型。采用随机数字表法将模型小鼠分为模型组,紫草素低(20 mg/kg)、中(30 mg/kg)、高(40 mg/kg)剂量组和阳性对照组(泼尼松龙,10 mg/kg),每组15只;另取15只作为正常对照组。各给药组大鼠按10 mL/kg灌胃相应药物,模型组和正常对照组灌胃等体积溶剂,每日1次,连续15 d。分别于给药后第5、10和15天观察小鼠搔抓行为;于给药后第7、15天评价小鼠皮损状况;末次给药结束后,HE染色观察小鼠皮损组织病理学变化;流式细胞术检测小鼠外周血中Th1/Th2比例;酶联免疫吸附试验(ELISA)检测小鼠血清中TSLP、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-4和干扰素(IFN)-γ水平;Western blot法检测小鼠皮损组织TSLP和OX40L蛋白表达水平。结果与正常对照组相比,模型组小鼠搔抓行为增加,皮肤炎性损伤加重;皮损组织中表皮和真皮区域均有密集的炎性细胞浸润,表皮增厚;外周血Th2细胞、TSLP、TNF-α和IL-4水平以及皮损组织TSLP和OX40L蛋白表达水平显著升高(P<0.05),外周血Th1细胞、Th1/Th2比例和IFN-γ水平显著下降(P<0.05)。与模型组相比,紫草素低、中、高剂量组小鼠第5、10、15天的搔抓行为减少,第7、15天皮肤炎性损伤依次减轻(P<0.05);小鼠表皮和真皮区域炎性细胞浸润和表皮增厚依次减轻,外周血Th2细胞、TSLP、TNF-α和IL-4水平以及皮损组织TSLP和OX40L蛋白表达水平依次降低(P<0.05),Th1细胞、Th1/Th2比例和IFN-γ水平依次升高(P<0.05)。结论紫草素可能通过抑制TSLP/OX40L通路维持AD小鼠外周血中Th1/Th2平衡,改善皮肤炎性损伤。Objective To investigate the effects of shikonin on thymic stromal lymphopoietin(TSLP)/OX40 ligand(OX40L)pathway and balance of helper T cell 1(Th1)/helper T cell 2(Th2)in mice with atopic dermatitis(AD).Methods AD mouse model was established by topical application of 2,4-dinitrofluorobenzene(DNFB).Model mice were randomly divided into model group,low shikonin group(10 mg/kg),medium shikonin group(20 mg/kg),high shikonin group(30 mg/kg),and the positive control group(prednisolone,10 mg/kg)by random number table method,with 15 mice in each group.Another 15 mice were used as the normal control group.The drug groups were given the corresponding drugs by gavage,with the volume of 10 mL/kg.The model group and the normal control group were given the same volume of solvent by gavage,once a day for 15 days.The scratch behavior of mice was observed on the 5th,10th and 15th day after administration.The condition of skin lesions was evaluated on the 7th and 15th days after administration.At the end of the last administration,hematoxylin eosin(HE)staining was used to observe the histopathological changes of skin lesions.The ratio of Th1/Th2 cells in peripheral blood of mice was detected by flow cytometry.Levels of TSLP,tumor necrosis factor(TNF)-α,interleukin(IL)-4 andγ-interferon(IFN-γ)were detected by enzyme linked immunosorbent assay(ELISA).Western blot assay was used to detect the protein expression levels of TSLP and OX40L.Results Compared with the normal control group,the scratching behavior increased and the skin inflammatory injury aggravated in model group.There were dense infiltration of inflammatory cells in the epidermis and dermis in the model group,and the epidermis was thickened.The blood levels of Th2 cells,TSLP,TNF-αand IL-4,and the protein expression levels of TSLP and OX40L in skin lesions were significantly higher(P<0.05),and the levels of Th1 cells,IFN-γand Th1/Th2 ratio were significantly lower(P<0.05).Compared with the model group,the scratching behavior were decreased on the 5th,the 10th and the

关 键 词：紫草素 皮炎 特应性 Th1-Th2平衡 OX40配体 胸腺基质淋巴细胞生成素/OX40L通路 

分 类 号：R275[医药卫生—中医皮科] R758.2[医药卫生—中医学]