| Objective To study whether Yuhuang decoction is effective in treating allergic contact dermatitis,then to explore the mechanism of action from the relationship between Th17 cell and Treg cell balance,and to judge the safety of Yuhuang decoction,to provide reliable and objective experimental basis for the wide application of Yuhuang decoction in clinic.Methods(1)validity: 32 mice were divided into blank control group,model control group,yuhuangtang group and drug control group(dexamethasone).The model of allergic contact dermatitis was established in the back of mice with 2,4 dinitrofluorobenzene(DNFB)repeatedly coated with model control group,yuhuangtang group and drug control group(dexamethasone),After sensitization,the corresponding drugs were given to each group,the changes of the skin lesions of the naked eye back of each group were observed,the histopathological changes were observed HE staining,the body weight and spleen weight of the mice were measured,and the spleen index was calculated.(2)Mechanism of action: The expression levels of IL-17、RORrt、IL-10 and Foxp3 genes and proteins in the back skin of each group of mice were detected by RT-PCR and Westen Blot methods,ELISA detection of serum IL-17、IL-10、IL-6 expression levels in each group.(3)skin long-term toxicity test :20 KM mice were divided into normal saline group and Yuhuangtang group.After back hair removal,each group of the back was given corresponding drugs for 14 days to observe the mental state of the mice in each group,whether there was death or not,and if there was death,dissect the mice.(4)skin irritation test :16 KM mice were divided into intact skin group and damaged skin group,and the control group was divided into Yuhuangtang group and normal saline group.The back groups were treated with the corresponding drugs for 14 days after the back depilation,and 1 h、24h and 48 h after removing the drugs were graded according to the order of skin irritation response.(5)Skin allergy test : The 18 KM mice were divided into Yuhuangtang group,saline group and positive control group.On the first day and the second day,the corresponding drugs were given in each group on the back of depilation.The positive control group was given 7%2,4dinitrofluorobenzene10 ul,and 1%2,4dinitrofluorobenzene 20 ul,immediately after the removal of the drug,24 h、48h、72h according to the skin allergy series score.Results(1)Effectiveness: The skin of the back of the blank control group was smooth,and the model control group obviously scabbed,mossy,and the back of the dexamethasone ointment group and the elm yellow group were all skin scabbed off,no moss,and a little hair was seen.The skin structure of the mice in the blank control group was clearly visible,the epidermis of the model control group was obviously thickened,the dermis lymphocytes increased,the epidermis of dexamethasone ointment group and elm yellow group became obviously thinner,and the dermis lymphocytes decreased.Compared with the mouse spleen index in the blank control group,the spleen index in the model control group was increased(P<0.01),and the spleen index in the dexamethasone ointment group and Yuhuangtang group was decreased compared with the model control group(P<0.01).(2)Mechanism of action: the gene and protein expression levels IL-17、RORγt the model control group were significantly higher than those IL-17 、RORγt in the blank control group(P<0.01).the gene and protein expression levels IL-17 、 ROR γ t the dexamethasone group and yuhuangtang group were significantly lower than those in the model control group(P<0.01).Genes and eggs IL-10、Foxp3 in the control group were compared with the expression levels of genes and proteins in the mice IL-10、Foxp3 in the control group the level of white expression was significantly decreased(P<0.01);compared with the gene and protein expression levels IL-10、Foxp3 the control group of the model,the gene and protein expression levels IL-10、Foxp3 the dexamethasone group and Yuhuangtang group were significantly increased(P<0.01).The serum IL-6 expression level of the model control group was significantly higher than that of the blank control group(P<0.01).Compared with the serum IL-6 expression level of the model control group,the expression level of serum IL6 in the dexamethasone group and Yuhuangtang group was decreased(P<0.05).The serum IL-17 expression levels were significantly the model control group was significantly higher than that of the blank control group(P<0.01),and the serum IL17 expression level of dexamethasone group and Yuhuangtang group was decreased compared with that of the model control group(P<0.01).The serum IL-10 expression level of the model control group was lower than that of the blank control group(P<0.05),and the serum IL10 IL-10 expression level of the model control group,the expression level of serum IL10 in the dexamethasone group and Yuhuangtang group was higher(P<0.05).(3)Dermal long-term toxicity experiment: during 14 days of medication,the diet,spirit,defecation and activity of the mice in the normal saline group and Yuhuang decoction group KM were normal,and no mice died during the period.There was no significant difference in weight gain between saline group and Yuhuangtang group.(4)Skin irritation experiment: the skin irritation response score of 1 h、24h and 48 h in the intact skin saline group and the intact skin Yuhuang decoction group was 0,the skin irritation response score of 1 h in the damaged skin saline group and the damaged skin Yuhuang decoction group was 2,and the skin irritation response score of 24 h and 48 was0.(5)Skin allergy experiment: the scores of immediate,24 h、48h、 and 72 h skin allergic reaction in Yuhuangtang group and physiological saline group were 0,and the sensitization rate was 0.The scores of immediate,24 h、48h、 and 72 h skin allergic reactions in the positive control group were 6.5,5.5,5.0,4.12,and the sensitization rate was100%.Conclusion:yuhuang decoction is used outside for allergic contact dermatitis has curative effect,and by intervention Th17/Treg cell balance to play a role,yuhuang decoction is non-toxic,skin irritant,skin allergic,has safety,can be widely used in clinical treatment. |
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