Study On The Effect Of Antipruritic Lotion On IL-4 And IFN-γ In Skin Lesions Of Atopic Dermatitis Model In Mic

Objective:The model of Atopic Dermatitis(AD)was set up by 2,4-dinitrochlorobenzene(DNCB)stimulating the skin of BALB/c mice.We investigated the therapeutic effect of Zhiyang Runfu cream on AD-like model of BALB/c mice by interfering JAK-STAT signaling pathway and its influence on the expression levels of interleukin-4(IL-4)and interferon-γ(IFN-γ)in mice skin lesions.Methods: Fifty SPF BALB/c mice were divided into 5 groups according to random number table,the group includes blank group,model group,mometasone furoate ointment group,delgocitinib ointment group and Zhiyang Runfu cream group,with 10 mice in each group.These groups were treated with back depilation.On the 1st-7th day of the experiment,100μl of 1%DNCB mixed liquid(olive oil: acetone = 1:4)was equably embrocated to the skin of the depilated part of the back of mice except the blank group.The purpose of this measure is stimulating AD-like reaction.From the2 nd week,100μl of 0.2%DNCB solution was used on the 3rd and 6th day of each week for intervention modeling and drug application to maintain the AD-like reaction.At the 15 th day of the experiment our model was steady.Mometasone furoate group was coated with mometasone furoate ointment(100μg once a day),delgocitinib ointment group was coated with delgocitinib ointment(100μg once a day),and Zhiyang Runfu cream group was coated with Zhiyang Runfu cream(100μg once a day)every day,while the blank group and model group were not treated.The process persisted for 14 days.These mice were killed on the 29 th day of the experiment and peeled back their AD lesions for pathological observation.The m RNA expression level and protein content of IL-4 and IFN-γ in AD lesions were determined by RT-q PCR method and western blot method respectively.Results: 1.After modeling,the number of scratches and the EASI score of skin lesions in the model group were apparently higher than the normal group(P<0.01).Following the effective cure,the AD lesions included Zhiyang Runfu cream 、 delgocitinib ointment and mometasone furoate ointment were meliorated,the scores of behavior observation tended to be normal,scratches times and the EASI score apparently declined(P<0.05).Under skin tissue microscope,the pathological conditions such as spinous hypertrophy,inflammatory cell infiltration and tissue edema were obviously ameliorate.2.RT-qPCR demonstrated that the expression of IL-4 m RNA in mice skin tissue increased and the expression of IFN-γ m RNA decreased after successfully establishing AD model.In contrast to model group,the expression of IL-4 m RNA in the Zhiyang Runfu cream,mometasone furoate ointment and delgocitinib ointment decreased(P<0.01),and the expression of IL-4 m RNA in the AD lesions of Zhiyang Runfu cream group was lower than that in the mometasone furoate ointment group(P<0.01).In contrast to model group,the levels of IFN-γ m RNA in the three treatment groups all increased(P<0.01,P<0.01,P<0.01),and the levels of IFN-γ m RNA in the Zhiyang Runfu cream group were apparently enhanced than mometasone furoate ointment group and delgocitinib ointment group(P<0.01,P<0.01).3.Western blot showed that DNCB stimulation could induce the contents of IL-4protein and inhibit the activation of IFN-γ protein.In clear contrast to model group,the IL-4 protein content in mometasone furoate ointment group,delgocitinib ointment group and Zhiyang Runfu cream group decreased apparently(P<0.01,P<0.01,P<0.01);Compared with mometasone furoate ointmen group,the content of IL-4 protein in Zhiyang Runfu cream group and delgocitinib ointment group decreased apparently(P<0.05,P<0.05).Following the effective cure,It is a legible contrast to the model group,the IFN-γ protein content in mometasone furoate ointment group,delgocitinib ointment group and Zhiyang Runfu cream group increased(P<0.01,P<0.01,P<0.01);The content of IFN-γ protein in Zhiyang Runfu cream group increased more obviously than that in delgocitinib ointment group and mometasone furoate ointment group(P<0.01,P<0.01)。Conclusion: The Zhiyang Runfu cream has obvious curative effect on AD-like symptoms induced by DNCB.And its mechanism probably achieved through the control of key cytokines IL-4 and IFN-γ in JAK-STAT signal transduction pathway,reducing the expression and content of IL-4 gene,and increasing the expression and content of IFN-γ gene to alleviate inflammatory reaction.