Effect of Anti-inflammation and Skin Hydration of AF-343 on Macrophage Raw 264.7 Cells and NC/Nga Mice with Atopic Dermatitis

Kim, Beom Joon; Lim, Yun Young; Kim, Hyeong Mi; Park, Eun Joo; Kim, Myeung Nam; Cho, Soo Muk; Choi, Chang Suk; Park, Ki Moon; Kim, Hyun Suck; Kim, Jong Keun; Hong, Yeon Pyo

Abstract

BACKGROUND
Inflammatory response on LPS and IFN-gamma induced Macrophage Raw 264.7 cells was secreted NO (nitric oxide) and PGE2 (prostaglandin E2) though expression of iNOS and COX-2. And many pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6 etc.) was secreted on LPS and IFN-gamma induced Macrophage Raw 264.7 cells, too. Atopy dermatitis was inflammatory skin disease with pruritus, xeroderma and specific eczema.
OBJECTIVE
We sought to effect of anti-inflammation and skin hydration of AF-343 on Macrophage Raw 264.7 cells and NC/Nga mice with Atopic Dermatitis.
METHODS
The immune response of Raw 264.7 cells were induced by LPS and IFN-gamma. Then LPS and IFN-gamma induced Raw 264.7 cells was measured NO, PGE2 production after treatment of different concentrations for AF-343. The related genes (iNOS, COX-2) for NO, PGE2 production were detected using Western blot in LPS and IFN-gamma induced Raw 264.7 cells after treatment of different concentrations for AF-343. Pro-inflammatory cytokines were detected, too. NC/Nga mice as Atopy dermatitis model was induced atopy dermatitis. Then NC/Nga mice with atopy dermatitis were performed oral administration of AF-343 for 1weeks. After oral administration of AF-343, TEWL was measured on skin tissues of NC/Nga mice with atopy dermatitis according to whether were performed oral administration of AF-343 or not. And pro-inflammatory cytokines and IgE was measured in serum, protein of skin tissues of NC/Nga mice. Skin tissues of NC/Nga mice were performed H&E staining, immunohistochemical staining for PCNA, Involucrin and filaggrin.
RESULTS
LPS and IFN-gamma induced Raw 264.7 cells was decreased NO, PGE2 production in dose-dependent after treatment of different concentrations for AF-343. The expression level of iNOS, COX-2 protein was decreased in dose-dependent, too. The related pro-inflammatory cytokines in media with LPS and IFN-gamma induced Raw 264.7 cells were decreased after treatment of different concentrations for AF-343. TEWL level of NC/Nga mice skin (back, ear) with atopy dermatitis according to whether were performed oral administration of AF-343 or not was decreased in NC/Nga mice with atopy dermatitis group was performed oral administration by AF-343. When NC/Nga mice group with atopy dermatitis was performed oral administration by AF-343, induced pro-inflammatory cytokines and IgE expression in serum, protein of back, ear skin tissues of each NC/Nga mice group was decreased. H&E stained Skin tissues of NC/Nga mice was confirmed that thickness of epidermis, dermis were decreased in NC/Nga mice group with atopy dermatitis was performed oral administration by AF-343 than NC/Nga mice group with atopy dermatitis. The expression of PCNA, involucrin and filaggrin were decreased in NC/Nga mice group with atopy dermatitis was performed oral administration by AF-343 than NC/Nga mice group with atopy dermatitis as results of immunihistochemical staining using specific antibodies such as PCNA as cell proliferation marker, involucrin and filaggrin as keratinocytes differentiation markers for skin tissues (back, ear) of NC/Nga mice.
CONCLUSION
We confirmed effect of anti-inflammation and skin hydration of AF-343 on Macrophage Raw 264.7 cells and NC/Nga mice with Atopic Dermatitis. In conclusion, AF-343 is expecting as therapeutics for atopic dermatitis.