[논문]화피(樺皮) 에탄올 추출물의 Ultraviolet B로 자극한 피부 각질 세포 보호 작용

최학순; 김현주; 이학송; 백승원; 김지은; 송용선

doi:10.13048/jkm.23020

화피(樺皮) 에탄올 추출물의 Ultraviolet B로 자극한 피부 각질 세포 보호 작용
Protective Effect of Betula Platyphylla on Ultraviolet B-irradiated HaCaT Keratinocytes 원문보기

Journal of Korean Medicine = 대한한의학회지, v.44 no.2, 2023년, pp.119 - 131

최학순 (원광대학교 한의학전문대학원 제3의학과) , 김현주 (원광대학교 한의학전문대학원 제3의학과) , 이학송 (원광대학교 한의과대학 한방재활의학교실) , 백승원 (원광대학교 한의과대학 한방재활의학교실) , 김지은 (원광대학교 한의과대학 한방재활의학교실) , 송용선 (원광대학교 한의학전문대학원 제3의학과)

Abstract ▼ AI-Helper

Objectives: Betula Platyphylla(BP) has been used as a analgesic, anti-microbial, anti-oxidant drug in Eastern Asia. However, it is still unknown whether BP ethanol extract could exhibit the inhibitory activities against ultraviolet B(UVB)-induced skin injury on human keratinocytes, HaCaT cells. This study was aimed to investigate the protective activity of BP ethanol extract on UVB-irradiated skin injury in HaCaT cells. Methods: The skin injury model of HaCaT cells was established under UVB stimulation. HaCaT keratinocyte cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB. Then, the cells were harvested to measure the cell viability, production of reactive oxygen species(ROS), pro-inflammatory cytokines such as interleukin(IL) 1-beta, IL-6, and tumor necrosis factor(TNF)-𝛼, hyaluronidase, type 1 collagen, matrix metalloproteinase(MMP)s. In addition, we examined the mitogen activated protein kinases(MAPKs) and inhibitory kappa B alpha(I𝜅;-B𝛼) as inhibitory mechanisms of BP ethanol extract. Results: The treatment of BP ethanol extract inhibited the UVBinduced cell death and ROS production in HaCaT cells. BP ethanol extract treatment inhibited the UVB-induced increase of IL-1beta, IL-6, and TNF-𝛼. BP ethanol extract treatment inhibited the increase of hyaluronidase, MMP and decrease of collagen. BP ethanol extract treatment inhibited the activation of MAPKs and the degradation of I𝜅-B𝛼. Conclusions: Our result suggest that treatment of BP ethanol extract could inhibit the UVB-induced skin injury via deactivation of MAPKs and nuclear factor kappa B(NF-𝜅B) in HaCaT cells. This study could suggest that BP ethanol extract could be a beneficial agent to prevent skin damage or inflammation.

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표/그림 (7)

그림 Fig. 1. The cytotoxicity of BP ethanol extract in human keratinocyte, HaCaT cells The cell viability was measured by MTT assay. HaCaT cells were incubated with or without BP ethanol extract as indicated doses for 24 h. The similar results were obtained from three additional experiments. *p < 0.05 significant as compared to normal.
그림 Fig. 2. The effects of BP ethanol extract on UVB-induced cell death in human keratinocyte, HaCaT cells The cell viability was measured by MTT assay. HaCaT cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB (200 mJ/cm2) for 24 h. The similar results were obtained from three additional experiments. *p < 0.05 significant as compared to normal. +p < 0.05 significant as compared to UVB alone
그림 Fig. 3. The effects of BP ethanol extract on UVB-induced ROS production in human keratinocyte, HaCaT cells The ROS production were measured by relative DCFDA intensity using FACS. HaCaT cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB for 24 h. The similar results were obtained from three additional experiments. *p < 0.05 significant as compared to normal. +p < 0.05 significant as compared to UVB alone.
그림 Fig. 4. The effects of BP ethanol extract on UVB-induced elevation of inflammatory cytokines in human keratinocyte, HaCaT cells The inflammatory cytokines were measured by realtime RT-PCR. HaCaT cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB for 24 h. The similar results were obtained from three additional experiments. *p < 0.05 significant as compared to normal. +p < 0.05 significant as compared to UVB alone.
그림 Fig. 5. The effects of BP ethanol extract on UVB-induced hyaluronidase 2 and 4 in human keratinocyte, HaCaT cells The mRNA expression of hyaluronidase 2 and 4 were measured by realtime RT-PCR. HaCaT cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB for 24 h. The similar results were obtained from three additional experiments. *p < 0.05 significant as compared to normal. +p < 0.05 significant as compared to UVB alone.
그림 Fig. 6. The effects of BP ethanol extract on UVB-induced collagen and MMP 1, and 9 in human keratinocyte, HaCaT cells The mRNA expression of MMP1 and 9 were measured by realtime RT-PCR. HaCaT cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB for 24 h. The similar results were obtained from three additional experiments. *p < 0.05 significant as compared to normal. +p < 0.05 significant as compared to UVB alone.
그림 Fig. 7. The Effects of BP ethanol extract on UVB-induced MAPKs activation and the degradation of Iκ-Bα in human keratinocyte, HaCaT cells The activation of MAPKs and degradation of Iκ-Bα were measured by Western blot. HaCaT cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB for 30 min. The similar results were obtained from three additional experiments.

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