Protective Effect of Betula Platyphylla on Ultraviolet B-irradiated HaCaT Keratinocytes

Hag Soon Choi; Hyun Joo Kim; Hark Song Lee; Seung Won Paik; Ji Eun Kim; Yung Sun Song; Choi, Hag Soon; Kim, Hyun Joo; Lee, Hark Song; Paik, Seung Won; Kim, Ji Eun; Song, Yung Sun

doi:10.13048/jkm.23020

JKM > Volume 44(2); 2023 > Article

Choi, Kim, Lee, Paik, Kim, and Song: Protective Effect of Betula Platyphylla on Ultraviolet B-irradiated HaCaT Keratinocytes

Original Article

The Journal of Korean Medicine 2023; 44(2): 119-131.

DOI: https://doi.org/10.13048/jkm.23020

Protective Effect of Betula Platyphylla on Ultraviolet B-irradiated HaCaT Keratinocytes

Hag Soon Choi¹

, Hyun Joo Kim¹

, Hark Song Lee²

, Seung Won Paik²

, Ji Eun Kim²

, Yung Sun Song¹^{, *}

¹Department of Third Medicine, Professional Graduate School of Korean Medicine, Wonkwang University

²Department of Korean Medicine Rehabilitation, College of Korean Medicine, Wonkwang University

Correspondence to: Yung Sun Song, Department of Third Medicine of Korean Medicine, Professional Graduate School of Oriental Medicine, Wonkwang University, 460 Iksan-daero, Iksan 54538, Korea, Tel: +82-63-270-1177, Fax: +82-63-270-1594, E-mail: yssong@wku.ac.kr

Received April 17, 2023 Revised May 6, 2023 Accepted May 10, 2023

Abstract

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.

Keywords: Betula Platyphylla(BP), ultraviolet B(UVB), keratinocytes, skin, inflammation

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.

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

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.

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.

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.

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.

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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