Construction Research of A Disease Syndrome Combination Mouse Model of Cold Dampness Syndrome with Atopic Dermatitis

WU Wenfeng; YU Yanxu; WU Qing; YE Siqi; ZHANG Yu; LIU Junfeng; MO Xiumei; CHEN Dacan; YAN Fenggen

Abstract：Objective To establish a mouse model of atopic dermatitis(AD)with cold dampness syndrome,and evaluate the model by detecting macro and micro indicators.Methods Balb/c mice were randomly divided into control group(CON),atopic dermatitis model group(AD),cold dampness syndrome model group(CDS),and a disease-syndrome combination model group(DSC),with 6 mice in each group.The mice in control group were treated with matrix liquid.In AD group,mice were treated with Dinitrochlorobenzene(DNCB)to simulate AD disease model.In CDS group,the mice were treated with matrix liquid,and combined with artificial climate box to mimic the cold-dampness environment[temperature(4±2)℃,relative humidity(90±5)%],and fed with high-fat feed to simulate the cold dampness syndrome.The mice in the DSC group were modeled by using a combination of DNCB and cold dampness syndrome simulation methods.The transepidermal water loss(TEWL),cold dampness syndrome,and scratching behavior of animals were measured.HE staining was used to observe the pathological changes of skin tissue on the back of mice,and the epidermal thickness and inflammatory cell infiltration were analyzed.Western blot was used to detect the expression levels of Claudin-1,TRPV3 and TRPV4.The ratio of Th17/Treg in lymph nodes was analyzed by flow cytometry.Results Compared with the AD group,the syndrome score of mice was significantly increased(P<0.001),the TEWL value and the epidermal thickness of the back skin of mice in the DSC group were significantly increased(P<0.05).The expression of Claudin-1 and TRPV3 proteins in the back skin decreased significantly(P<0.01),while the expression of TRPV4 protein increased significantly(P<0.05)in mice of the DSC group compared with those in the AD group.Conclusion Using DNCB to establish the model of AD,combined with the cold dampness environment and high-fat diet,the mouse model of AD with cold dampness syndrome can be successfully established.This study provides an ideal animal model for further exploring the establishment of animal model of AD combined with cold dampness syndrome,and the efficacy of TCM compound on AD syndrome mouse model.