Anti-inflammatory Effects of Myrrh Ethanol Extract on Particulate Matter-induced Skin Injury

Young Hee Jung; Yeun Wha Roh; Myongsoo Chong; Jung, Young Hee; Roh, Yeun Wha; Chong, Myongsoo

doi:10.13048/jkm.22026

JKM > Volume 43(3); 2022 > Article

Jung, Roh, and Chong: Anti-inflammatory Effects of Myrrh Ethanol Extract on Particulate Matter-induced Skin Injury

Original Article

The Journal of Korean Medicine 2022; 43(3): 1-15.

Published online: September 1, 2022

DOI: https://doi.org/10.13048/jkm.22026

Anti-inflammatory Effects of Myrrh Ethanol Extract on Particulate Matter-induced Skin Injury

Young Hee Jung¹

, Yeun Wha Roh²

, Myongsoo Chong³^{, *}

¹Dept. of Third Medicine, Professional Graduate School of Korean Medicine, Wonkwang University

²Dept. of Korean Pathology, College of Korean Medicine, Wonkwang University

³Dept. of Preventive Medicine, College of Korean Medicine & Research Center of Traditional Korean Medicine, Wonkwang University

Correspondence to: Myongsoo Chong, 460 Wonkwang University Oriental Medicine School, Iksan-daero, Iksan-si, Jeollabuk-do, Tel: +82-63-850-6912, Fax: +82-63-852-5594, E-mail: neurokid@wku.ac.kr

Received March 31, 2022 Revised May 27, 2022 Accepted July 22, 2022

Abstract

Objectives

Myrrh have been used as a traditional remedy to treat infectious and inflammatory diseases. However, it is largely unknown whether myrrh ethanol extract could exhibit the inhibitory activities against particulate matter (PM)-induced skin injury on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the inhibitory activity of myrrh ethanol extract on PM-induced skin injury in HaCaT cells.

Methods

To investigate the inhibitory effects of myrrh ethanol extract in HaCaT cells, the skin injury model of HaCaT cells was established under PM treatment. HaCaT keratinocyte cells were pre-treated with myrrh ethanol extract for 1 h, and then stimulated with PM. Then, the cells were harvested to measure the cell viability, reactive oxygen species (ROS), pro-inflammatory cytokines including interleukin (IL) 1-beta, IL-6, and tumor necrosis factor (TNF)-α, hyaluronidase, collagen, MMPs. In addition, we examined the mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha (Iκ-Bα) as inhibitory mechanisms of myrrh ethanol extract.

Results

The treatment of myrrh ethanol extract inhibited the PM-induced cell death and ROS production in HaCaT cells. In addition, myrrh ethanol extract treatment inhibited the PM-induced elevation of IL-1beta, IL-6, and TNF-α . Also, myrrh ethanol extract treatment inhibited the increase of hyaluronidase, MMP and decrease of collagen. Furthermore, myrrh ethanol extract treatment inhibited the activation of MAPKs and the degradation of Iκ-Bα.

Conclusions

Our result suggest that treatment of myrrh ethanol extract could inhibit the PM-induced skin injury via deactivation of MAPKs and nuclear factor (NF)-κB in HaCaT cells. This study could suggest that myrrh ethanol extract could be a beneficial agent to prevent skin damage or inflammation.

Keywords: Myrrh, Myrrh ethanol extract, Particulate matter, Keratinocyte, Skin, Inflammation

Fig. 1

The cytotoxicity of Myrrh ethanol extract in HaCaT cells

The cell viability was measured by MTT assay. HaCaT cells were incubated with or without Myrrh 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 cytotoxicity of PM (particulate matter) in HaCaT cells

The cell viability was measured by MTT assay. HaCaT cells were incubated with or without PM as indicated doses for 24 h. The similar results were obtained from three additional experiments. ^*p < 0.05 significant as compared to normal. N.S. indicates non-significant.

Fig. 3

The effects of Myrrh ethanol extract on PM-induced cell death in HaCaT cells

The cell viability was measured by MTT assay. HaCaT cells were pre-treated with Myrrh ethanol extract for 1 h, and then incubated with PM as indicated doses 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 PM alone

Fig. 4

The effects of PM (particulate matter) on ROS production in HaCaT cells

The ROS production were measured by relative DCFDA intensity using FACS. HaCaT cells were incubated with or without PM as indicated doses for 24 h. The similar results were obtained from three additional experiments. ^*p < 0.05 significant as compared to normal.

Fig. 5

The effects of Myrrh ethanol extract on PM-induced ROS production in HaCaT cells

The ROS production were measured by relative DCFDA intensity using FACS. HaCaT cells were pre-treated with Myrrh ethanol extract for 1 h, and then incubated with PM as indicated doses 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 PM alone

Fig. 6

The effects of PM (particulate matter) on pro-inflammatory cytokines in HaCaT cells

The pro-inflammatory cytokines were measured by realtime RT-PCR. HaCaT cells were incubated with or without PM as indicated doses for 24 h. The similar results were obtained from three additional experiments. ^*p < 0.05 significant as compared to normal.

Fig. 7

The effects of Myrrh ethanol extract on PM-induced pro-inflammatory cytokines in HaCaT cells

The pro-inflammatory cytokines were measured by realtime RT-PCR. HaCaT cells were pre-treated with Myrrh ethanol extract for 1 h, and then incubated with PM as indicated doses 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 PM alone

Fig. 8

The effects of Myrrh ethanol extract on PM-induced hyaluronidase 2 and 4 in HaCaT cells

The mRNA expression of hyaluronidase 2 and 4 were measured by realtime RT-PCR. HaCaT cells were pre-treated with Myrrh ethanol extract for 1 h, and then incubated with PM as indicated doses 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 PM alone

Fig. 9

The effects of Myrrh ethanol extract on PM-induced collagen and MMP 2 and 9 in HaCaT cells

The mRNA expression of COL1A1, MMP-2 and MMP-9 were measured by realtime RT-PCR. HaCaT cells were pre-treated with Myrrh ethanol extract for 1 h, and then incubated with PM as indicated doses 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 PM alone

Fig. 10

The Effects of Myrrh ethanol extract on PM-induced MAPKs and the degradation of Iκ-Bα in HaCaT cells

HaCaT cells were pre-treated with Myrrh ethanol extract for 1 h, and then incubated with PM for 30 min. The phosphorylation of ERK1/2, P38 and JNK, and the degradation of Iκ-Bα were analyzed by western blotting. Total ERK1/2, P38, JNK and β-actin were used as loading controls. The similar results were obtained from three additional experiments.

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