Your browser does not support the NLM PubReader view.

Go to this page to see a list of supporting browsers.

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

Article information

J Korean Med. 2022;43(3):1-15
Publication date (electronic) : 2022 September 1
doi : https://doi.org/10.13048/jkm.22026
Young Hee Jung1orcid_icon, Yeun Wha Roh2orcid_icon, Myongsoo Chong3,orcid_icon
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 2022 March 31; Revised 2022 May 27; Accepted 2022 July 22.

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.

References

1. Yun M.S., Kim C., Hwang J.K.. 2019; Agastache rugosa Kuntze attenuates UVB-induced photoaging in hairless mice through the regulation of MAPK/AP-1 and TGF-β/Smad pathways. J Microbiol Biotechnol 29:1349–1360. https://dx.doi.org/10.4014/jmb.1908.08020 .
2. Jo I.J.. 2022;Inhibitory activity of Terminalia chebula extract against TNF-α/IFN-γ-induced chemokine increase on human keratinocyte, HaCaT cells. Kor J Herbol 37(3):41–47. http://dx.doi.org/10.6116/kjh.2022.37.3.41 .
3. Kwon D.J., Bae Y.S., Ju S.M., Goh A.R., Choi S.Y., Park J.. 2011;Casuarinin suppresses TNF-α-induced ICAM-1 expression via blockade of NF-κB activation in HaCaT cells. Biochem Biophys Res Commun 409:780–785. https://dx.doi.org/10.1016/j.bbrc.2011.05.088 .
4. Lee Y.J., Seo S.H.. 2021;The effects of perception of particulate matter harmfulness on skin health management behavior. J Kor Soc Cosmetol 27(5):1229–1237. https://doi.org/10.52660/JKSC.2021.27.5.1229 .
5. Pan T.L., Wang P.W., Aljuffali I.A., Huang C.T., Lee C.W., Fang J.Y.. 2015;The impact of urban particulate pollution on skin barrier function and the subsequent drug absorption. J Derma Sci 78:51–60. https://dx.doi.org/10.1016/j.jdermsci.2015.01.011 .
6. Hu R., Xie X.Y., Xu S.K., Wang Y.N., Jiang M., Wen L.R., et al. 2017;PM2.5 Exposure elicits oxidative stress responses and mitochondrial apoptosis pathway activation in HaCaT keratinocytes. Chin Med J 130:2205–2214. https://dx.doi.org/10.4103/0366-6999.212942 .
7. Choe Y., Moon K.M., Yoo J.C., Byun J., Hwang S., Moon D.K., Woo D.K.. 2021;Atorvastatin and fluvastatin can reduce IL-1β-induced inflammatory responses in human keratinocytes. J Life Science 31(4):418–424. https://doi.org/10.5352/JLS.2021.31.4.418 .
8. Herbology Textbook Compilation Committee. 2020. Herbal Medicine 4th Edth ed. Seoul: Youngrimsa. p. 442–443.
9. Baek S.J., Kim D.H.. 2016;The Study on Anti-obesity of Myrrh ethanol extract. Kor J Herbol 31(4):11–18. http://dx.doi.org/10.6116/kjh.2016.31.4.11 .
10. Salama A., Ibrahim W., El-Nimr T., Abd El M.A., Tousson E.. 2014;Effect of myrrh extract (Mirazid? on experimentally diabetic rats. Pharmacologia 5(4):135–142. https://dx.doi.org/10.5567/pharmacologia.2014.135.142 .
11. Kim D.C., Yoon C.S., Ko W., Lee D.S., Kim D.S., Cho H.K., et al. 2015;Anti-inflammatory Effects of 1β, 6α-Dihydroxyeudesm-4(15)-ene Isolated from Myrrh on LPS-induced Neuroinflammation in BV2 cells. Kor J Pharmacogn 46(1):12–16.
12. Baek H.S., Kang S.K., Auh Q.S., Chun Y.H., Hong J.P.. 2013;Effect of antibacterial effects of myrrh, rhatany, chamomomilla. J Oral Medicine and Pain 38(4):299–312. http://dx.doi.org/10.14476/jomp.2013.38.4.299 .
13. El-Shahat M., El-Abd S., Alkafafy M., El-Khatib G.. 2012;Potential chemoprevention of diethylnitrosamine-induced hepatocarcinogenesis in rats: myrrh (Commiphora molmol) vs. turmeric (Curcuma longa). Acta Histochemica 114(5):421–428. https://dx.doi.org/10.1016/j.acthis.2011.08.002 .
14. Ahn J., Kim D.S., Cho H.K., Kim Y.C., Kim S.Y., Oh H., et al. 2015;Ameliorating effects of HPM-1 on Scopolamine-induced memory impairments in mice. Kor J Pharmacogn. 46(3):243–249.
15. Yang S.J., Kim K.M., Song J.W., Lee S.H.. 2021;Relaxing effect of novel cosmetic ingredient using Lactobacillus gasseri HDB1102 on skin problems caused by particulate matter. J Kor Soc Cosmetol 27(5):1152–1158. https://dx.doi.org/10.52660/jksc.2021.27.5.1152 .
16. Seo S.K., Ku C.S., Son B.M., Lee S.G., Kang E.M., Kim M.O., et al. 2016;Anti-pollution capacity of mixtures of Anemarrhena asphodeloides root extract, β -glucan and poly-γ-glutamic acid from stimulation with particulate matter. J Invest Cosmetol 12(4):313–322. https://dx.doi.org/10.5352/JLS.2020.30.2.191 .
17. Jang A.S.. 2014;Impact of particulate matter on health. J Kor Med Assoc 57(9):763–768. http://dx.doi.org/10.5124/jkma.2014.57.9.763 .
18. Lee J., Kim H., Kim J., Han C.J.. 2019;A study on the awareness of skin harmfulness and skin reaction symptoms and skin care behavior about the particulate matter of adult women. J Kor Soc Cosmetol 25(6):1309–1315.
19. Park U.K., Oh J.H., Park S.I., Lim J.H., Kwon S.J., Kim E.J., et al. 2020;Antimicrobial activities of Commiphora myrrha Holmes mixture extract against Clavibacter michiganensis subspmichiganesis and capsici . Natural Science 31:23–33.
20. Chen Y., Zhou C., Ge Z., Liu Y., Liu Y., Feng W., et al. 2013;Composition and potential anticancer activities of essential oils obtained from myrrh and frankincense. Oncol Lett 6(4):1140–1146. https://dx.doi.org/10.3892/ol.2013.1520 .
21. Kim M.S., Bae G.S., Park K.C., Koo B.S., Kim B.J., Lee H.J., et al. 2012;Myrrh inhibits LPS-induced inflammatory response and protects from cecal ligation and puncture -induced sepsis. Evid Based Complement Alternat Med 2012:278718. https://dx.doi.org/10.1155/2012/278718 .
22. Poon F., Kang S., Chien A.L.. 2015;Mechanisms and treatments of photoaging. Photodermatol Photoimmunol Photomed 31:65–74. https://dx.doi.org/10.1111/phpp.12145 .
23. Lee W.R., Kim K.H., An H.J., Kim J.Y., Han S.M., Lee K.G., et al. 2014;The effects of bee venom on tumor necrosis factor (TNF)-α induced inflammatory human HaCaT keratinocytes. Kor J Pharmacogn 45(3):256–261.
24. Yoo H.R., Lee C.H.. 2018;A change in sebum, moisture and satisfaction according to hyaluronic acid content. Kor J Oral Maxillofacial Pathology 42(3):73–86. https://doi.org/10.17779/KAOMP.2018.42.3.003 .
25. Myung D.B., Han H.S., Shin J.S., Park J.Y., Hwang H.J., Kim H.J., et al. 2019;Hydrangenol isolated from the leaves of Hydrangea serrata attenuates wrinkle formation and repairs skin moisture in UVB-irradiated hairless mice. Nutrients 11(10):2354. https://dx.doi.org/10.3390/nu11102354 .
26. Kim E., Hwang K., Lee J., Han S.Y., Kim E.M., Park J., et al. 2018;Skin protective effect of Epigallocatechin gallate. Int J Mol Sci 19(1):173. https://dx.doi.org/10.3390/ijms19010173 .
27. Kang S.J., Choi B.R., Kim S.H., Yi H.Y., Park H.R., Park S.J., et al. 2015;Inhibitory effects of pomegranate concentrated solution on the activities of hyaluronidase, tyrosinase, and metalloproteinase. J Cosmet Sci. 66(3):145–159.
28. Yun M.E., Lee Y.S., Lee Y.J., Park Y.M., Park S.N.. 2018;Antimicrobial, antioxidant and cellular protective effects of Houttuynia cordata extract and fraction. Appl Chem Eng 29:452–460. https://doi.org/10.14478/ace.2018.1035 .
29. Pittayapruek P., Meephansan J., Prapapan O., Komine M., Ohtsuki M.. 2016;Role of matrix metalloproteinases in photoaging and photocarcinogenesis. Int J Mol Sci. 17:868–888. https://doi.org/10.3390/ijms17060868 .
30. Zhang M., Zhou J., Wang L., Li B., Guo J., Guan X., et al. 2014;Caffeic acid reduces cutaneous tumor necrosis factor alpha (TNF-α), IL-6 and IL-1β levels and ameliorates skin edema in acute and chronic model of cutaneous inflammation in mice. Biol Pharm Bull 37:347–354. https://dx.doi.org/10.1248/bpb.b13-00459 .
31. Bae G.S., Park S.J.. 2019;Anti-inflammatory Effect of Nypa fruticans Wurmb on tumor necrosis factor (TNF)-α-induced inflammatory response in HaCaT cells. Kor J Herbol 34(1):51–57. https://doi.org/10.6116/kjh.2019.34.1.51 .
32. Choi H.J., Alam M.B., Baek M.E., Kwon Y.G., Lim J.Y., Lee S.H.. 2020;Protection against UVB-induced photoaging by Nypa fruticans via inhibition of MAPK/ AP-1/MMP-1 signaling. Oxid Med Cell Longev 2020:290536. https://dx.doi.org/10.1155/2020/2905362 .
33. Liu A., Zhao W., Zhang B., Tu Y., Wang Q., Li J.. 2020;Cimifugin ameliorates imiquimod-induced psoriasis by inhibiting oxidative stress and inflammation via NF-κ B/MAPK pathway. Biosci Rep 40(6):BSR20200471. https://dx.doi.org/10.1042/BSR20200471 .

Article information Continued

Fig. 1

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

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

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

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

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

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

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

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

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

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.