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白鮮皮 추출물이 DNCB로 유발된 NC/Nga 생쥐의 아토피 피부염에 미치는 영향

The Effects of Dictamni Radicis Cortex Extracts on DNCB Induced Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse

김윤희|한재경|이윤실
대한한방소아과학회 (The Association of Pediatrics of Korean Medicine)|2014.11|대한한방소아과학회지 (The Journal of Pediatrics of Korean Medicine)|제28권 제4호|pp. 1-29

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키워드

Atopic dermatitisDNCBNC/Nga mouseDictamni Radicis CortexMC/9 murine mast cell
주제분류한의학

초록

ObjectivesDictamni Radicis Cortex extracts (DRC) has been known to suppress allergic reaction, however the cellulartarget of DRC and its mode of action remain unclear. The purpose of this study is to investigate the effects ofDictamni Radicis Cortex extracts on DNCB induced atopic dermatitis-like skin lesions of NC/Nga mouse. MethodsThis study was designed to investigate the effects of DRC extract in the DNP-IgE-induced activation of MC/9murine mast cell lines in vitro and in the DNCB-induced activation of NC/Nga mouse in vivo. For thisinvestigation, We examined IL-4, IL-5, IL-6, IL-13, TNF-α and GM-CSF mRNA expression by Real-time PCR,IL-13, MIP-1α production by ELISA analysis and manifestations of NFAT1, NFAT2, AP-1 and NF-κB p65transcription factors by western blotting in vitro. Then, we examined WBC, eosinophil and neutrophil in NC/Ngamouse, IL-5, IL-13 in serum, IFN-γ, IL-4 in the spleenocyte culture supernatant, the absolute cell numbers ofCD4+, CD8+, +Gr-1+CD11b, B220+CD23+ in the ALN, PBMCs and dorsal skin, IL-5, IL-13 in the dorsal skin byReal-time PCR and the distribution of mast cells by H&E and toluidine blue. ResultsIn vitro the mRNA expression of IL-4, IL-5, IL-6, IL-13, TNF-α, GM-CSF and IL-13, MIP-1α production byELISA analysis were completely abolished by DRC and the western blot analysis decreased the expression of mastcell-specific transcription factors including NFAT-1, NF-κB p65. In vivo DRC oral adminstration also decreasedthe counts of WBC, eosinophils and inflammatory cytokines such as IL-13 and IgE in the serum. DRC oraladminstration elevated IL-4 level in the spleenocyte culture supernatant. DRC oral adminstration decreased totalALN cells, total skin cells, cell numbers of CD4+, B220+CD23+ in the ALN, +Gr-1+CD11b in the PBMCs andCD4+, CD8+ in the dorsal skin. The mRNA expression of IL-5, IL-13, thickness of epidermis, inflammationimmune cells and mast cells were abolished by DRC in the dorsal skin. ConclusionsHistological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Ngamouse were much improved by DRC oral adminstration. These results, therefore, suggest that DRC can regulatemolecular mediators and immune cells that are functionally associated with atopic dermatitis induced in NC/Ngamouse, and may play an important role in recovering AD symptoms.

Abstract

ObjectivesDictamni Radicis Cortex extracts (DRC) has been known to suppress allergic reaction, however the cellulartarget of DRC and its mode of action remain unclear. The purpose of this study is to investigate the effects ofDictamni Radicis Cortex extracts on DNCB induced atopic dermatitis-like skin lesions of NC/Nga mouse. MethodsThis study was designed to investigate the effects of DRC extract in the DNP-IgE-induced activation of MC/9murine mast cell lines in vitro and in the DNCB-induced activation of NC/Nga mouse in vivo. For thisinvestigation, We examined IL-4, IL-5, IL-6, IL-13, TNF-α and GM-CSF mRNA expression by Real-time PCR,IL-13, MIP-1α production by ELISA analysis and manifestations of NFAT1, NFAT2, AP-1 and NF-κB p65transcription factors by western blotting in vitro. Then, we examined WBC, eosinophil and neutrophil in NC/Ngamouse, IL-5, IL-13 in serum, IFN-γ, IL-4 in the spleenocyte culture supernatant, the absolute cell numbers ofCD4+, CD8+, +Gr-1+CD11b, B220+CD23+ in the ALN, PBMCs and dorsal skin, IL-5, IL-13 in the dorsal skin byReal-time PCR and the distribution of mast cells by H&E and toluidine blue. ResultsIn vitro the mRNA expression of IL-4, IL-5, IL-6, IL-13, TNF-α, GM-CSF and IL-13, MIP-1α production byELISA analysis were completely abolished by DRC and the western blot analysis decreased the expression of mastcell-specific transcription factors including NFAT-1, NF-κB p65. In vivo DRC oral adminstration also decreasedthe counts of WBC, eosinophils and inflammatory cytokines such as IL-13 and IgE in the serum. DRC oraladminstration elevated IL-4 level in the spleenocyte culture supernatant. DRC oral adminstration decreased totalALN cells, total skin cells, cell numbers of CD4+, B220+CD23+ in the ALN, +Gr-1+CD11b in the PBMCs andCD4+, CD8+ in the dorsal skin. The mRNA expression of IL-5, IL-13, thickness of epidermis, inflammationimmune cells and mast cells were abolished by DRC in the dorsal skin. ConclusionsHistological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Ngamouse were much improved by DRC oral adminstration. These results, therefore, suggest that DRC can regulatemolecular mediators and immune cells that are functionally associated with atopic dermatitis induced in NC/Ngamouse, and may play an important role in recovering AD symptoms.

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