The Expression Of AQP3 In Keratinocyte And The Expression Of Serum IL-17?IL-23 In Atopic Dermatitis And The Correction Of Them

Objective:To investigate the relationship of the skin barrier function and immune disorders,we establish atopic dermatitis mice model and test the expression of aquaporin3 in keratinocyte and the serum IL-17?IL-23.Methods:51 female BALB/c mice,which are 6 weeks old.They were divided into three groups,normal group(n=17)?model group(n=17)and treatment group(n=17).A week before the experiment,we removed the mice back hair using 8%sodium sulfide starch solution.The area of removed hair is 3.0cm×2.0cm.Establishing the normal control group:Firstly,we smeared 200ul of acetone/oil(3:1)mixture on the mice back which had been removed hair.The eighth day,we smeared 200?l of acetone/oil(3:1)mixture on the mice back(1/3days)and the total times was 13.The 26th day,we injected normal saline solution in enterocoelia(1/3days).The 42th day,we narcoticzed the mice.Model group:Firstly,we smeared 1%2,4dinitrofluorobenzene(DNCB)of acetone/oil(3:1)mixture(dose:200?l)on the mice back.The eighth day,we smeared 0.2%DNCB of acetone/oil(3:1)mixture(dose:200?l)on the mice back(1/3days)and the total times was 13.The 26th day,we injected normal saline solution in enterocoelia(1/3 days).The 42th day,we narcotized the mice.Treatment mice group:The first day of the experiment,we smeared 1%2,4dinitrofluorobenzene(DNCB)of acetone/oil(3:1)mixture(dose:200?l)on the mice back.The eighth day,we smeared 0.2%DNCB of acetone/oil(3:1)mixture(dose:200?l)on the mice back(1/3days)and the total times was 13.The 26th day,we injected interferony in enterocoelia(1/3 days).The 42th day,we narcotized the mice.We narcotized mice and get the blood,putting it in EP under 37?for 20min and centrifuged 20minutes.We measured serum IgE?IL-17?IL-23.We picked up the skin of three mice groups under aseptic conditions,then we observe the pathological change after HE staining of the skin tissue.We observe AQP3 in three mice groups through Western blot.Results:1.The score of mice skin inflammation:The score of AD group(9.00)is higher than that of the control group(0.00),the score of treatment group(4.00)is lower than AD group(9.00),the differences were statistically significant(P<0.05).2.The result of histopathological test in three groups:The control group was not observed inflammatory cells infiltration.We can observe thickening of stratum spinosum in AD mice skin under the microscopic observation.The skin lesion of the treatment group was lighter than model group3.The result of Western Blot:We compared the ratio of AQP3/?actin in three groups.The ratio of AD group is(0.76±0.13)higher than that of control group(0.49±0.12),The ratio of treatment group is group(0.68±0.09)is lower than that of AD group(0.76±0.13,the differences were statistically significant(P<0.05).The spearman correlation of the ratio of AQP3/?actin and AD was rs=0.726,the difference was statistically significant(P<0.01).The pearson correlation of the ratio of AQP3/pactin and IgE?IL-17?IL-23 was r=0.780?r=0.640?r=0.746.The differences were statistically signifycant(P<0.01).4.The result of ELISA:We compared the expression of IgE in three groups,the total IgE expression level of AD group(158.94±3.67)is higher than that of control group(20.21±3.08),the total IgE expression level of treatment group(143.79±8.31)is lower than that of AD group(158.94±3.67),The differences were statistically significant(P<0.05).The IL-17 expression level of AD group(32.74±2.57)is higher than that of control group(17.11±3.35),The IL-17 expression level of treatment group(19.80±2.88)is lower than that of AD group(32.74±2.57),the differences were statistically significant(P<0.05).The IL-23 expression level of AD group(98.08±3.18)is higher than that of control group(49.57±2.63),The IL-23 expression level of treatment group(62.50±2.83)is lower than that of AD group(98.08±3.18),the differences were statistically significant(P<0.05).The spearman correlation coefficient of the IgE?IL-17?IL-23 expression level and the degree of the AD was rs=0.713?rs=0.625?rs=0.697,respectively.The differences were statistically significant(P<0.01).The pearson correlation of IgE and IL-17?IL-23 was r=0.630?r=0.756.The differences were statistically significant(P<0.01).Conclusions:1.The expression level of the serum IL-17,IL-23 is associated with AD.2.The expression level of AQP3 in KC is associated with AD.3.The skin barrier disfunction and the immune disorder of AD influence each other.