| Objective:To detect the expression of antioxidant nuclearfactor erythroid-2 related factor 2(Nrf2),NF-?B p65 and keratin(Krt)10,14,17 in skin lesions BALB/c famle mice model of atopic dermatitis(AD).To explore the relationships between the three and study the mechanism of Nrf2 improve dermatitis AD.To clear the feasibility of the AD animal models and provide further research direction by antioxidant intervention treatment of AD.Methods:In this study,40 BLAB/c female mice of SPF grade with about 6 weeks around old and 20g around were selected and randomly divided into such 4 groups as normal control group,the AD model group,treatment control group and treatment group.The methods of sensitizing and stimulating AD model:AD group,treatment control group and treatment group to adapt to the environment in the laboratory to start building after 1 week,On the oth,7th and 14th days of the experiment,0.1%OVA solution intraperitoneal injection in mice abdomen.In the 14th day of the experiment,the back of the mice was depilated,On the 21st day,taking appropriate thickness of the size of the gauze dipped in 0.1%OVA solution put in spot test apparatus,and covered in AD mice back hair removal area,fixed,applied frequency is 3 times per week,a total of three weeks.Modeling:normal control group before each dose,time with three groups(solution for fresh vial aluminum hydroxide gel),before daub dose,time with three groups(solution for PBS).On the 42nd day,the treatment group was givensulforaphane(SFN),the treatment control group treated jojoba oil for one hour,than all were given 0.1%OVA.The AD model group was given 0.1%OVA;Normal control group was treated with PBS,1 time/day,7 days in total.On the 49th day,we observed the number of scratches in the four groups of mice.The weight and spleen weight of the four groups were recorded.The concentrations of total IgE and TNF-? concentrations were detected by ELISA,and histopathological observation was carried out with HE staining,and the expression of Nrf2,NF-?B p65 and Krt10,14 and 17 in mouse skin was detected by Western blot.Results:1.The mice skin inflammation degree grade:AD group compared with the normal group,treatment control group compared with the treatment of control group were statistically significant difference(P<0.01).2.The number of spleen index and scratching results:AD group?treatment control group and treatment group mice spleen index and scratching frequencyare all higher than the normal control group(P<0.05).3.Results of histopathological examination:there was no damage in the back skin of the normal control group,no abnormal structural changes were observed under the microscope,and no inflammatory cells were infiltrated.AD and treatment control group visible epidermis,dermis thickening of different level,there is a lot of chronic inflammatory cells infiltration.The skin thickening of the treatment group was lower than the control treatment group.4.Elisa results:Compared with the normal group,the total IgE and TNF-? concentration of the AD group,the treatment group and the treatment control group are respectively increased(P<0.05).5.Western Blot test:(1)the expression of Nrf2:the Nrf2/GAPDH ratio was not statistically significant different between the four groups.The expression of Nrf2 nucleoprotein:the relative expression of Nrf2 in the AD group are lower than that in the normal control group,while the treatment group was significantly higher than the control treatment group(P<0.05).(2)The expression of keratin:the ratios of Krt10/GAPDH and Krtl4/GAPDH of the AD group are higher than normal group,and the treatment group significantly decreased compared with the treatment control group(P<0.05).The expression of Krt17 AD of the other 3 groups were higher than the normal group,and the treatment group was higher than the treatment control group(P<0.05).(3)The expression of NF-?B p65:the ratios of the total NF-rcB p65/GAPDH of the four groups were not statistically significant different.The expression of NF-?B p65:the NF-?B p65 of the AD group obviously increased compared with normal group;while the treatment group was lower than the treatment control group(P<0.05).Conclusion:1.Using OVA method to establish AD animal model,AD dermatitis was improved with the SFN treatment.2.The external Nrf2 agonist SFN effectively improved the expression of Nrf2,and verified that Nrf2 could be used to treat AD dermatitis.3.The external Nrf2 agonist SFN can down-regulate the expression of the inflammatory factor NF-?B p65,and regulate the expression of Krt10,14 and 17.It is suggested that the antioxidant Nrf2 may be treated by down-regulating inflammatory factors and regulating epidermal proliferation and differentiation mechanism. |
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