研究目的
To explore the effect of Fn14 inhibition on experimental atopic dermatitis and the effect of TWEAK on keratinocytes in the atopic inflammatory microenvironment.
研究成果
Fn14 deficiency ameliorates skin lesion in the murine model of atopic dermatitis, possibly through suppressing the production of inflammatory cytokines and altering the expression profile of TNF receptors, leading to less apoptosis of keratinocytes. Targeting the TWEAK/Fn14 pathway may be a promising therapeutic approach for patients with atopic dermatitis.
研究不足
The study did not elucidate the effect of TWEAK on the barrier function of skin in atopic dermatitis. The in-vitro model used may not fully replicate the complexity of atopic dermatitis in humans.
1：Experimental Design and Method Selection:
The study utilized both in-vivo and in-vitro models to explore the role of TWEAK/Fn14 signaling in atopic dermatitis. In-vivo, atopic dermatitis was induced in wild-type and Fn14 knock-out BALB/c mice using a 2,4-dinitrofluorobenzene (DNFB) method. In-vitro, human primary keratinocytes were cultured under atopic dermatitis-like inflammation conditions.
2：Sample Selection and Data Sources:
Skin tissue and sera were collected from patients with atopic dermatitis and healthy donors. Male BALB/c mice aged 8 weeks were used for the in-vivo model.
3：List of Experimental Equipment and Materials:
Equipment included the ABI PRISM 7900HT Sequence Detection System for qRT-PCR, Axioskop II fluorescence microscope for immunofluorescence, and electrophoresis gels for Western blotting. Materials included DNFB for inducing atopic dermatitis, TWEAK for stimulation, and various antibodies for detection.
4：Experimental Procedures and Operational Workflow:
The in-vivo model involved sensitization and maintenance of inflammation with DNFB. The in-vitro model involved stimulating keratinocytes with an inflammatory cocktail. Immunohistochemistry, immunofluorescence, qRT-PCR, ELISA, and Western blotting were performed to analyze the effects.
5：Data Analysis Methods:
Data were analyzed using GraphPad Prism version 5.0, with ANOVA for comparing more than two groups and a two-tailed Student’s t-test for comparing two groups.
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