The Expression And Significance Of MiR-146a In Patients With Atopic Dermatitis And Atopic Dermatitis-like Mice Model

Objective:1.To detect the expression of Thl(CD4+IFN-γ+)、Th2(CD4+IL-4+)cells in atopic dermatitis-like model mice spleen and miR-146a in mice skin lesions.2.To study the effect of PTQX particles on the expression of Thl、Th2 cells in atopic dermatitis-like model mice spleen and miR-146a in mice skin lesions.3.To study the expression of Thl,Th2 in the peripheral blood mononuclear cell(PBMC)and miR-146a in the serum of atopic dermatitis patients,and to explore the correlation between miR-146a expression and SCORAD,IGA score and different syndrome types of atopic dermatitis patients.Methods:1.Experimental study Male C57BL/6 mice were randomly divided into control group,model group,PTQX particle group.Use DNFB induce to establish atopic dermatitis-like model.Using a shaving razor and depilatory cream to remove the hair of mice back.The area size about 2cmX2cm.On the first day,50ul 0.5%DNFB solution which configured by acetone and olive oil was applied to the control group,PTQX particle group mice back skin.Since the fifth day,the mice right ear and back respectively applied 20ul and 50ul 0.2%DNFB solution every 3 days(5,8,11,14 day).The control group mice was applied the same amount of acetone and olive oil(acetone and olive oil without damage to mice skin).PTQX particle group mice had medication from the first day until the fourteenth day.After modeled,the mice were observed back,ear skin lesions,ear swelling degree and skin pathology.Flow cytometry was used to detect the expression of Thl and Th2 cells in mice spleen.RT-PCR was used to detect the expression of miR-146a in mice skin lesions.2.Clinical study 25 AD patients which in accord with Hanifin and Rajika AD diagnostic criteria and IGA score more than 3 points were included,16 healthy people were included.The patients were scored according to the SCORAD score.Referring to the expert consensus of TCM diagnosis and treatment program of AD in 2013 for syndrome differentiation.Detected IgE expression and allergen in patients.Flow cytometry was used to detect the expression of Thl and Th2 in PBMC.RT-PCR was used to detect the expression of miR-146a in serum.To explore the correlation between miR-146a expression and SCORAD,IGA score.The expression of miR-146a in different syndrome types was analyzed.Results:1.The control group mice back and ear skin were normal.The model group mice back and ear skin were thickened,rough,erythema,desquamation,crusting.The PTQX particle group mice back and ear skin were mildly thickened,rough,pale erythema,a little desquamation,crusting.The control group mice ear were normal,The model group mice ear swelling significantly,The PTQX particle group mice ear were mildly swelling,three groups compared with each other were statistical significance.The control group mice skin pathology were normal.In the model group,the pathological features of the skin were epidermal parakeratosis,hyperkeratosis,spongy edema,spinous layer thickening,and a large number of lymphocytic infiltration in the dermis.In the PTQX particle group,the pathological features of the skin showed epidermis mild parakeratosis,hyperkeratosis,spongy edema,slight spinous layer thickening,and a small amount of lymphocytes infiltration in the dermis.2.The Thl cells in model group and PTQX particle group were lower than that in control group,the difference are not statistically significant(P>0.05).The Th2 in model group was higher than that in control group and PTQX particle group,the difference are statistically significant(P<0.01).The Th2 in PTQX particle group was higher than that in control group,the difference is not statistically significant(P>0.05).The Th1/Th2 in model group was lower than that in control group,the difference is statistically significant(P<0.01).The Th1/Th2 in model group was lower than that in PTQX particle group,the difference is statistically significant(P<0.05).The Th1/Th2 in PTQX particle group was lower than that in control group,the difference is not statistically significant(P>0.05).3.The miR-146a in model group was higher than that in control group,the difference is statistically significant(P<0.01).The miR-146a in PTQX particle group was higher than that in control group,the difference is statistically significant(P<0.05).The miR-146a in PTQX particle group was lower than that in control group,the difference is not statistically significant(P>0.05).4.Among the 25 AD patients,6 cases with heart and spleen heat retention syndrome,12 cases with heart fire and spleen deficiency syndrome,4 cases with damp aggregate and spleen deficiency syndrome,3 cases with blood deficiency and wind-dryness syndrome.The miR-146a expression in patients with heart fire and spleen deficiency syndrome is the highest than other three groups,the difference are not statistically significant(P>0.05).5.The Thl cells in AD group was lower than that in control group,the difference is statistically significant(P<0.01).The Thl cells in AD group was higher than that in control group,the difference is not statistically significant(P>0.05).The Thl/Th2 in AD group was lower than that in control group,the difference is statistically significant(P<0.05).6.The miR-146a expression in AD group was higher than that in control group,the difference is statistically significant(P<0.05).The miR-146a expression were not correlation with SCORAD and IGA score.Conclusions:1.Atopic dermatitis-like C57BL/6 mice model induced by DNFB is an ideal model to study the pathogenesis of atopic dermatitis.2.PTQX particle can inhibit the inflammatory response of atopic dermatitis,its mechanism may be through regulating Thl/Th2 immune imbalance and inhibiting miR-146a expression.3.MiR-146a may be involved in the pathogenesis of atopic dermatitis.