Your browser does not support the NLM PubReader view.

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

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

Article information

J Korean Med. 2023;44(2):119-131
Hag Soon Choi1orcid_icon, Hyun Joo Kim1orcid_icon, Hark Song Lee2orcid_icon, Seung Won Paik2orcid_icon, Ji Eun Kim2orcid_icon, Yung Sun Song1,orcid_icon
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 2023 April 17; Revised 2023 May 6; Accepted 2023 May 10.

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.

References

1. Choi EY, Sohn HY, Lee JT. 2019;The Antioxidant and Anti-aging Effects of Treatment with Schisandra chinensis Seed Fractions in UVB-irradiated Human HaCaT cells. J Life Sci 29(10):1071–9. https://doi.org/10.5352/JLS.2019.29.10.1071.
2. Alexandra AO, Betty Y, John AD. 2014;Ultraviolet radiation, aging and skin: prevention of damage by tropical cAMP manipulation. Molecules 19:6202–19. https://doi.org/10.3390/molecules19056202.
3. Edwin DL. 2016;Skin aging and oxidative stress: Equol’s anti-aging effects via biochemical and molecular mechanisms. Ageing Res Rev 31:36–54. https://doi.org/10.1016/j.arr.2016.08.001.
4. Masamitsu I, Hideya A, Masaki Y, Toko N, Mary M. 2009;Photoaging of the skin. J Anti Aging Med 6:46–59. https://doi.org/10.3793/jaam.6.46.
5. Martin GM, Austad SN, Johson TE. 1996;Genetic analysis of aging: role of oxidative damage and environmental stresses. Nat Genet 13(1):25–34. https://doi.org/10.1038/ng0596-25.
6. Pittayapruek P, Meephansan J, Prapapan O, Komine M, Ohtsuki M. 2016;Role of matrix metalloproteinases in photoaging and photocarcinogenesis. Int J Mol Sci 17:868–88. https://doi.org/10.3390/ijms17060868.
7. Kim SM, Chung JH. 2008;Berberine prevents UV-induced MMP-1 and reduction of type | procollagen expression in human dermal fibroblasts. Phytomedicine 15(9):749–53. https://doi.org/10.1016/j.phymed.2007.11.004.
8. Chae YM. 1996. Jungyakyakmyeongsajun Beijing: Chinese Medicine Publisher. p. 44.
9. Sin MG, Jeong BY. 2003. Dohae Hyangyakdaesajeon Seoul: Yeongrimsa. p. 803–4.
10. Sin MG. 2002. Clinical Traditional Herbalogy Seoul: Yeongrimsa. p. 399–400. p. 844.
11. Kim YJ, Lee JD, Lee YH. 2007;The Effects of Betula Platyphylla on Cartilage Protecion, Anti-inflammatory and Analgesic Activity in Arthritis. The Joumal of Korean Acupunct & Moxibustion Society 2007;24(2):73–81.
12. Kim WI, Kim JE, Lee SH, Moon YS, Lee SH, Park SY, Na CS. 2013;Antioxidative and Antimicrobial Activities of Water- and Ethanol-Extracts from Betula platyphylla var. japonica, Punica granatum and Rhus javanica. Kor J Herbology 28(3):45–51. https://doi.org/10.6116/kjh.2013.28.3.45.
13. Lee CH, Cho YM, Park ES, Shin CS, Lee JY, Jeong HS. 2009;In vivo Immune Activity of Sap of the White Birch(Betula platyphylla var. japonica). Kor J Food Sci Technol 41(4):413–6.
14. Park SY, Na CS, Jeong WC, Lee JC. 2012;A Literature Study of Pericarpium Granati and Cortex Betulae Platyphyllae. The Journal of Korean Oriental Medical Ophthalmology & Otolaryngology & Dermatology 25(3):13–33. https://doi.org/10.6114/jkood.2012.25.3.013.
15. Herbal Pharmacology Textbook Compilation Committee. 1992. Herbal Pharmacology Seoul: Shinil Publisher. p. 469–71.
16. The National College of Oriental Medicine Herbology Classroom. 2008. Herbology Seoul: Youngrimsa. p. 190–1,728.
17. An JB. 2002. Dictionary of oriental medicine Mock Yang.
18. Huh JE, Hopng JM, Baek HY, Lee JD, Choi DY, Park SS. 2011;Anti-inflammatory and anti-nociceptive effect of Betula platyphylla var. japonica in human interleukin-1β-stimulated fibroblast-like synoviocytes and in experimental animal models. J Ethnopharmacol 135(1):126–34. https://doi.org/10.1016/j.jep.2011.03.005.
19. Ryu MH, Park EK, Kim YH, Lee YA, Lee SH, Hong SJ, Baek YH, Park DS, Han JS, Yoo MC, Yang HI, Kim KS. 2006;Anti-inflammatory effects of an ethanolic extract from Betula Platyphylla. J Korean Oriental Med 27(1):184–95.
20. Schwiebs A, Radeke HH. 2018;Immunopharmacological Activity of Betulin in Inflammation-associated Carcinogenesis. Anticancer Agents Med Chem 18(5):645–51. http://dx.doi.org/10.2174/1871520617666171012124820.
21. Lim JP. 2011;Antibacterial activity against S. mutans or P. gingivalis and anti-inflammatory effect of betulae cortex. Korean J. Oriental Physiology & Pathology 25(4):635–40.
22. Matsuda H, Ishikado A, Nishida N, Ninomiya K, Fujiwara H, Kobayashi Y, Yoshikawa M. 1998;Hepatoprotective, superoxide scavenging, and antioxidative activities of aromatic constituents from the bark of Betula Platyphylla var. Japonica. Bioorganic & Medicinal Chemistry letters 8:2939–44. https://doi.org/10.1016/s0960-894x(98)00528-9.
23. Joo SY. 2013;Antioxidant activities of medicinal plant extracts. J Korean Soc Food Sci Nutr 42(4):512–9. https://doi.org/10.3746/jkfn.2013.42.4.512.
24. Ju EM, Lee SE, Hwang HJ, Kim JH. 2004;Antioxidant and anticancer activity of exteact from Betula platyphylla var. japonica. Life Sciences 74:1013–26. https://doi.org/10.1016/j.lfs.2003.07.025.
25. Halliday GM. 2005;Inflammation, gene mutation and photoimmunosuppression in response to UVR induced oxidative damage contributes to photocarcinogenesis. Mutation Re search 2005;571(1–2):107–20. https://doi.org/10.1016/j.mrfmmm.2004.09.013.
26. Trouba KJ, Hamadeh HK, Amin RP, Germolec DR. 2002;Oxidative stress and its role in skin disease. Antioxid. Redox. Signal 4:665–73. https://doi.org/10.1089/15230860260220175.
27. Ansel J, Luger TA, Lowry D. 1988;The expression and modulation of IL-1 alpha in murine keratinocytes. J Immunol 140:2274–8.
28. Diniyah N, Alam MB, Choi HJ, Lee SH. 2020;Lablab Purpureus Protects HaCaT Cells from Oxidative Stress-Induced Cell Death through Nrf2-Mediated Heme Oxygenase-1 Expression via the Activation of p38 and ERK1/2. Int J Mol Sci 21(22):8583. https://doi.org/10.3390/ijms21228583.
29. Gates T. 2007;Atopic dermatitis : diagnosis, treatment, and aeromedical implications. Aviat Space Environ Med 78:29–37.
30. Verdier-Sévrain S, Bonté F. 2007;Skin hydration: a review on its molecular mechanisms. J Cosmet Dermatol 6(2):75–82. https://doi.org/10.1111/j.1473-2165.2007.00300.x.
31. Myung DB, Han HS, Shin JS, Park JY, Hwang HJ, Kim HJ, Ahn HS, Lee SH, Lee KT. 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://doi.org/10.3390/nu11102354.
32. Kang SJ, Choi BR, Kim SH, Yi HY, Park HR, Park SJ, Song CH, Park JH, Lee YJ, Kwang S. 2015;Inhibitory effects of pomegranate concentrated solution on the activities of hyaluronidase, tyrosinase, and metalloproteinase. J Cosmet Sci 66(3):145–59.
33. Scharffetter K, Wlaschek M, Hogg A, Bolsen K, Schothorst A, Goerz G, Krieg T, Plewig G. 1991;UVA irradiation induces collagenase in human dermal fibroblasts in vitro and in vivo. Arch. Dermatol. Res 283(8):506. https://doi.org/10.1007/bf00371923.
34. Brenneisen P, Wenk J, Klotz LO, Wlaschek M, Briviba K, Krieg T, Sies H, Scharffetter-Kochanek K. 1998;Central role of ferrous/ferric iron in the ultraviolet B irradiation -mediated signaling pathway leading to increased interstitial collagenase (matrix-degrading metalloproteinase (MMP)-1) and stromelysin-1 (MMP-3) mRNA levels in cultured human dermal fibroblasts. J Biol Chem 273(9):5279–87. https://doi.org/10.1074/jbc.273.9.5279.
35. Zhang M, Zhou J, Wang L, Li B, Guo J, Guan X, Han Q, Zhang H. 2014;Caffeic acid reduces cutaneous tumor necrosis factor alpha (TNF-α), IL-6 and IL-1 levels and ameliorates skin edema in acute and chronic model of cutaneous inflammation in mice. Biol. Pharm. Bull 37:347–54. https://doi.org/10.1248/bpb.b13-00459.
36. Celec P. 2004;Nuclear factor kappa b-molecular biomedicine: the next generation. Biomed. Pharmacother 58:365–71.
37. Qi T, Huan Y, Enmie L, Hya W. 2017;P38/ERK MAPK signaling pathways are involved in the regulation of filaggrin and involucrin by IL-17. Mol, Med. Rep 16(6):8863–7. https://doi.org/10.3892%2Fmmr.2017.7689.
38. Choi HJ, Alam MB, Baek ME, Kwon YG, Lim JY, Lee SH. 2020;Protection against UVB-Induced Photoaging by Nypa fruticans via Inhibition of MAPK/AP-1/MMP-1 Signaling. Oxid Med Cell Longev 2020:2905362. https://doi.org/10.1155/2020/2905362.
39. 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-κB/MAPK pathway. Biosci Rep 40(6):BSR20200471. https://doi.org/10.1042%2FBSR20200471.

Article information Continued

Fig. 1

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

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

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

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

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

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

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.