镇心安神方对特应性皮炎样动物模型维生素D及丝聚蛋白表达的影响

Effect of Zhenxin Anshen Recipe on Expression of Vitamin D and Filaggrin of Atopic Dermatitis-like Mice Model

目的探讨镇心安神方(ZXASF)对特应性皮炎(AD)样小鼠模型血清及皮肤组织1,25(OH)2-D3、维生素D受体及Filaggrin表达的影响。方法采用1-氯-2,4-二硝基苯(DNCB)反复刺激BALB/c小鼠背部皮肤,建立AD样动物模型。54只小鼠随机分为6组(正常对照组、模型组、维生素D组、ZXASF低、中、高剂量组),每组9只。造模14 d后,ZXASF低、中、高剂量组分别按含ZXASF生药量9.9 g/(kg·d)、18.18 g/(kg·d)及36.36 g/(kg·d)的颗粒水溶液灌胃,维生素D组灌服维生素D滴剂2000 IU/(kg·d),模型组及正常对照组灌服生理盐水0.1 mL/(10g·d),共治疗14 d,观察各组模型小鼠皮损面积及皮炎评分,ELISA检测模型动物血清1,25(OH)2-D3、维生素D受体及皮肤组织Filaggrin含量,HE染色观察皮损组织病理学变化,免疫组织化学检测皮肤组织FLG的表达。结果末次给药后模型组皮炎评分、皮炎面积均高于正常对照组(P<0.01),模型组皮肤组织病理学表现为角化过度、棘层肥厚、海绵形成、真皮层炎性细胞浸润,ZXASF低、中、高剂量组及维生素D组小鼠背部皮损的红斑、糜烂、渗液及结痂减轻,皮炎评分及皮损面积均低于模型组(P<0.05)。与正常对照组比较,模型组血清1,25(OH)2D3、维生素D受体及皮肤组织Filaggrin含量均降低(P<0.05)。与模型组比较,ZXASF中剂量组血清1,25(OH)2D3、维生素D受体及皮肤组织Filaggrin含量升高(P<0.05),ZXASF低、高剂量组血清1,25(OH)2D3含量低于维生素D组(P<0.05),ZXASF中剂量组血清VDR含量高于维生素D组(P<0.05)。结论ZXASF可以改善AD样动物模型皮损,上调动物模型血清1,25(OH)2-D3、维生素D受体及皮肤组织Filaggrin的表达,以中剂量组作用最为显著。

Objective To explore the effects of Zhenxin Anshen(ZXAS)Recipe on expression of 1,25(OH)2-D3,Vitamin D receptor(VDR)and Filaggrin(FLG)in serum and skin lesions of atopic dermatitis-like mice model.Methods The animal model of atopic dermatitis was established by repeatedly applied on the shaved dorsal skin of BALB/c mice with 1-chloro-2-dinitrobenzene(DNCB).Fifty-four mice were randomly divided into 6 groups,which were normal control group,model group,Vit D group,low-dose(LD),medium-dose(MD),and high-dose(HD)ZXAS groups respectively,9 mice in each group.After 14 days of model being established,in the LD,MD,and HD ZXAS groups,9.9 g/(kg·d),18.18 g/(kg·d),and 36.36 g/(kg·d)were used for gavage respectively.In the Vit D group,2000 IU/(kg·d)were used for gavage with Vitamin D drops.The normal control group and the model group were given by 0.1 mL/(10 g·d)of saline.All groups were treated for 14 days.Dermatitis score and skin lesions area in atopic dermatitis model were observed.ELISA was used to detect the expression of 1,25(OH)2-D3,VDR and FLG in serum and skin lesions.The histopathological changes were observed by HE staining.Immunohistochemistry was used for expression of FLG in skin.Results After the last administration,compared with the normal control group,dermatitis score and dermatitis area in the model group increased(P<0.01).Histopathological results of skin tissue showed that hyperkeratosis,acanthosis,spongiform edema,inflammatory cell infiltration in the dermis.Compared with the model group,symptoms of erythema,erosions,scabs were relieved.Dermatitis score and skin lesions area of LD,MD,HD groups and Vit D group were lower than model group(P<0.05).Compared with the normal control group,the content of 1,25(OH)2-D3,VDR in serum and the expression of filaggrin in skin lesions in the model group decreased(P<0.05).Compared with the model group,the content of 1,25(OH)2-D3,VDR in serum and the expression of filaggrin in skin lesions increased in the MD ZXAS group(P<0.05).Compared with the Vit D group,the content of 1,25(OH)2-D3 in serum of LD and HD groups decreased(P<0.05),and the content of VDR in serum of MD group increased(P<0.05).Conclusion ZXAS Recipe can improve the skin lesions of AD-like animal model,and up-regulate the expression of 1,25(OH)2-D3,VDR and FLG in serum of AD-like mice model,especially in the middle dose group.