Horse Oil Mitigates Oxidative Damage to Human HaCaT Keratinocytes Caused by Ultraviolet B Irradiation

Piao, Mei Jing,Kang, Kyoung Ah,Zhen, Ao Xuan,Kang, Hee Kyoung,Koh, Young Sang,Kim, Bong Seok,Hyun, Jin Won MDPI 2019 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.20 No.6

<P>Horse oil products have been used in skin care for a long time in traditional medicine, but the biological effects of horse oil on the skin remain unclear. This study was conducted to evaluate the protective effect of horse oil on ultraviolet B (UVB)-induced oxidative stress in human HaCaT keratinocytes. Horse oil significantly reduced UVB-induced intracellular reactive oxygen species and intracellular oxidative damage to lipids, proteins, and DNA. Horse oil absorbed light in the UVB range of the electromagnetic spectrum and suppressed the generation of cyclobutane pyrimidine dimers, a photoproduct of UVB irradiation. Western blotting showed that horse oil increased the UVB-induced Bcl-2/Bax ratio, inhibited mitochondria-mediated apoptosis and matrix metalloproteinase expression, and altered mitogen-activated protein kinase signaling-related proteins. These effects were conferred by increased phosphorylation of extracellular signal-regulated kinase 1/2 and decreased phosphorylation of p38 and c-Jun N-terminal kinase 1/2. Additionally, horse oil reduced UVB-induced binding of activator protein 1 to the matrix metalloproteinase-1 promoter site. These results indicate that horse oil protects human HaCaT keratinocytes from UVB-induced oxidative stress by absorbing UVB radiation and removing reactive oxygen species, thereby protecting cells from structural damage and preventing cell death and aging. In conclusion, horse oil is a potential skin protectant against skin damage involving oxidative stress.</P>

Protective Effect of the Ethyl Acetate Fraction of <i>Sargassum muticum</i> Against Ultraviolet B–Irradiated Damage in Human Keratinocytes

Piao, Mei Jing,Yoon, Weon Jong,Kang, Hee Kyoung,Yoo, Eun Sook,Koh, Young Sang,Kim, Dong Sam,Lee, Nam Ho,Hyun, Jin Won Molecular Diversity Preservation International (MD 2011 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.12 No.11

<P>The aim of this study was to investigate the cytoprotective properties of the ethyl acetate fraction of <I>Sargassum muticum</I> (SME) against ultraviolet B (UVB)-induced cell damage in human keratinocytes (HaCaT cells). SME exhibited scavenging activity toward the 1,1-diphenyl-2-picrylhydrazyl radicals and hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>) and UVB-induced intracellular reactive oxygen species (ROS). SME also scavenged the hydroxyl radicals generated by the Fenton reaction (FeSO<SUB>4</SUB> + H<SUB>2</SUB>O<SUB>2</SUB>), which was detected using electron spin resonance spectrometry. In addition, SME decreased the level of lipid peroxidation that was increased by UVB radiation, and restored the level of protein expression and the activities of antioxidant enzymes that were decreased by UVB radiation. Furthermore, SME reduced UVB-induced apoptosis as shown by decreased DNA fragmentation and numbers of apoptotic bodies. These results suggest that SME protects human keratinocytes against UVB-induced oxidative stress by enhancing antioxidant activity in cells, thereby inhibiting apoptosis.</P>

Protective Effect of Fisetin (3,7,3`,4`-Tetrahydroxyflavone) against γ-Irradiation-Induced Oxidative Stress and Cell Damage

( Mei Jing Piao ),( Ki Cheon Kim ),( Sung Wook Chae ),( Young Sam Keum ),( Hye Sun Kim ),( Jin Won Hyun ) 한국응용약물학회 2013 Biomolecules & Therapeutics(구 응용약물학회지) Vol.21 No.3

Ionizing radiation can induce cellular oxidative stress through the generation of reactive oxygen species, resulting in cell damage and cell death. The aim of this study was to determine whether the antioxidant effects of the fl avonoid fi setin (3,7,3`,4`-tetrahydroxyfl avone) included the radioprotection of cells exposed to γ-irradiation. Fisetin reduced the levels of intracellular reactive oxygen species generated by γ-irradiation and thereby protected cells against γ-irradiation-induced membrane lipid peroxidation, DNA damage, and protein carbonylation. In addition, fi setin maintained the viability of irradiated cells by partially inhibiting γ-irradiation-induced apoptosis and restoring mitochondrial membrane potential. These effects suggest that the cellular protective effects of fi setin against γ-irradiation are mainly due to its inhibition of reactive oxygen species generation.

Diphlorethohydroxycarmalol Suppresses Ultraviolet B-Induced Matrix Metalloproteinases via Inhibition of JNK and ERK Signaling in Human Keratinocytes

( Mei Jing Piao ),( Madduma Hewage Susara Ruwan Kumara ),( Ki Cheon Kim ),( Kyoung Ah Kang ),( Hee Kyoung Kang ),( Nam Ho Lee ),( Jin Won Hyun ) 한국응용약물학회 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.6

Skin aging is the most readily observable process involved in human aging. Ultraviolet B (UVB) radiation causes photo-oxidation via generation of reactive oxygen species (ROS), thereby damaging the nucleus and cytoplasm of skin cells and ultimately leading to cell death. Recent studies have shown that high levels of solar UVB irradiation induce the synthesis of matrix metalloproteinases (MMPs) in skin fibroblasts, causing photo-aging and tumor progression. The MMP family is involved in the breakdown of extracellular matrix in normal physiological processes such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes such as arthritis and metastasis. We investigated the effect of diphlorethohydroxycarmalol (DPHC) against damage induced by UVB radiation in human skin keratinocytes. In UVB-irradiated cells, DPHC significantly reduced expression of MMP mRNA and protein, as well as activation of MMPs. Furthermore, DPHC reduced phosphorylation of ERK and JNK, which act upstream of c-Fos and c-Jun, respectively; consequently, DPHC inhibited the expression of c-Fos and c-Jun, which are key components of activator protein-1 (AP-1, up-regulator of MMPs). Additionally, DPHC abolished the DNA-binding activity of AP-1, and thereby prevented AP-1-mediated transcriptional activation. These data demonstrate that by inactivating ERK and JNK, DPHC inhibits induction of MMPs triggered by UVB radiation.

Phloroglucinol inhibits ultraviolet B radiation-induced oxidative stress in the mouse skin

Piao, Mei Jing,Ahn, Mee Jung,Kang, Kyoung Ah,Kim, Ki Cheon,Zheng, Jian,Yao, Cheng Wen,Cha, Ji Won,Hyun, Chang Lim,Kang, Hee Kyoung,Lee, Nam Ho,Hyun, Jin Won Informa Healthcare 2014 International journal of radiation biology Vol.90 No.10

<P><I>Purpose</I>: Previously we demonstrated that phloroglucinol (1,3,5-trihydroxybenzene) protected human HaCaT keratinocytes against ultraviolet B (UVB, 280-320 nm)-induced oxidative stress <I>in vitro</I> by scavenging intracellular reactive oxygen species (ROS). The current study investigated whether phloroglucinol could similarly protect the mouse skin against UVB-induced oxidative tissue damage <I>in vivo</I>.</P><P><I>Materials and methods</I>: Male 7-week-old Balb/c mice were divided into the following untreated normal control, phloroglucinol only-treated, vehicle plus UVB (30 or 60 mJ/cm<SUP>2</SUP>)-exposed, and phloroglucinol (10 or 50 mg/ml) plus UVB (30 or 60 mJ/cm<SUP>2</SUP>)-treated groups. Following UVB exposure, phloroglucinol or phosphate buffered saline vehicle was applied to the dorsal skin of each mouse daily for 3 days. Studies were conducted at 24 h after the last of the UVB exposures. Histopathological analyses of dorsal skin lesions were performed on all mice. In addition, the levels of UVB-provoked injury to cellular components, including DNA, proteins, and lipids were detected by levels of 8-oxoguanine (8-oxoG), protein carbonyls, and 8-isoprostane. Apoptosis were assessed by using western blot for B-cell lymphoma-2-associated X protein (Bax) and activated caspase-3 expression, by using immunohistochemistry.</P><P><I>Results</I>: UVB radiation increased the thickness of the epidermis and the dermis, and also stimulated the accumulation of mast cells in the irradiated skin. However, treatment with phloroglucinol significantly decreased all of these parameters. Furthermore, phloroglucinol decreased UVB-provoked injury to cellular components, including DNA, proteins, and lipids; down-regulated the expression of phospho-histone H2A.X in the injured skin; and reduced the UVB-generated levels of 8-oxoG, protein carbonyls, and 8-isoprostane, which are all markers of oxidative stress. In addition, phloroglucinol attenuated the UVB-induced expression of the pro-apoptotic proteins, Bax protein, and activated caspase-3.</P><P><I>Conclusion</I>: These results suggest that phloroglucinol safeguards the mouse skin against UVB-induced oxidative stress and apoptosis.</P>

Antioxidant Effects of the Ethanol Extract from Flower of <i>Camellia japonica</i> via Scavenging of Reactive Oxygen Species and Induction of Antioxidant Enzymes

Piao, Mei Jing,Yoo, Eun Sook,Koh, Young Sang,Kang, Hee Kyoung,Kim, Junoh,Kim, Yong Jin,Kang, Hak Hee,Hyun, Jin Won Molecular Diversity Preservation International (MD 2011 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.12 No.4

<P>The aim of this study was to investigate the antioxidant properties of the ethanol extract of the flower of <I>Camellia japonica</I> (<I>Camellia</I> extract). <I>Camellia</I> extract exhibited 1,1-diphenyl-2-picrylhydrazyl radical and intracellular reactive oxygen species (ROS) scavenging activity in human HaCaT keratinocytes. In addition, <I>Camellia</I> extract scavenged superoxide anion generated by xanthine/xanthine oxidase and hydroxyl radical generated by the Fenton reaction (FeSO<SUB>4</SUB> + H<SUB>2</SUB>O<SUB>2</SUB>) in a cell-free system, which was detected by electron spin resonance spectrometry. Furthermore, <I>Camellia</I> extract increased the protein expressions and activity of cellular antioxidant enzymes, such as superoxide dismutase, catalase and glutathione peroxidase. These results suggest that <I>Camellia</I> extract exhibits antioxidant properties by scavenging ROS and enhancing antioxidant enzymes. <I>Camellia</I> extract contained quercetin, quercetin-3-<I>O</I>-glucoside, quercitrin and kaempferol, which are antioxidant compounds.</P>

Phloroglucinol Attenuates Ultraviolet B-Induced 8-Oxoguanine Formation in Human HaCaT Keratinocytes through Akt and Erk- Mediated Nrf2/Ogg1 Signaling Pathways

( Mei Jing Piao ),( Ki Cheon Kim ),( Kyoung Ah Kang ),( Pincha Devage Sameera Madushan Fernando ),( Herath Mudiyanselage Udari Lakmini Herath ),( Jin Won Hyun ) 한국응용약물학회 2021 Biomolecules & Therapeutics(구 응용약물학회지) Vol.29 No.1

Ultraviolet B (UVB) radiation causes DNA base modifications. One of these changes leads to the generation of 8-oxoguanine (8- oxoG) due to oxidative stress. In human skin, this modification may induce sunburn, inflammation, and aging and may ultimately result in cancer. We investigated whether phloroglucinol (1,3,5-trihydroxybenzene), by enhancing the expression and activity of 8-oxoG DNA glycosylase 1 (Ogg1), had an effect on the capacity of UVB-exposed human HaCaT keratinocytes to repair oxidative DNA damage. Here, the effects of phloroglucinol were investigated using a luciferase activity assay, reverse transcription-polymerase chain reactions, western blot analysis, and a chromatin immunoprecipitation assay. Phloroglucinol restored Ogg1 activity and decreased the formation of 8-oxoG in UVB-exposed cells. Moreover, phloroglucinol increased Ogg1 transcription and protein expression, counteracting the UVB-induced reduction in Ogg1 levels. Phloroglucinol also enhanced the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) as well as Nrf2 binding to an antioxidant response element located in the Ogg1 gene promoter. UVB exposure inhibited the phosphorylation of protein kinase B (PKB or Akt) and extracellular signal-regulated kinase (Erk), two major enzymes involved in cell protection against oxidative stress, regulating the activity of Nrf2. Akt and Erk phosphorylation was restored by phloroglucinol in the UVB-exposed keratinocytes. These results indicated that phloroglucinol attenuated UVB-induced 8-oxoG formation in keratinocytes via an Akt/Erk-dependent, Nrf2/Ogg1-mediated signaling pathway.

The ethyl acetate fraction of <i>Sargassum muticum</i> attenuates ultraviolet B radiation-induced apoptotic cell death via regulation of MAPK- and caspase-dependent signaling pathways in human HaCaT keratinocytes

Piao, Mei Jing,Kim, Ki Cheon,Zheng, Jian,Yao, Cheng Wen,Cha, Ji Won,Boo, Sun Jin,Yoon, Weon Jong,Kang, Hee Kyoung,Yoo, Eun Sook,Koh, Young Sang,Ko, Mi Hee,Lee, Nam Ho,Hyun, Jin Won Informa Healthcare USA, Inc. 2014 PHARMACEUTICAL BIOLOGY Vol.52 No.9

<P><I>Context</I>: Our previous work demonstrated that an ethyl acetate extract derived from <I>Sargassum muticum</I> (Yendo) Fenshol (SME) protected human HaCaT keratinocytes against ultraviolet B (UVB)-induced oxidative stress by increasing antioxidant activity in the cells, thereby inhibiting apoptosis.</P><P><I>Objective</I>: The aim of the current study was to further elucidate the anti-apoptotic mechanism of SME against UVB-induced cell damage.</P><P><I>Materials and methods</I>: The expression levels of several apoptotic-associated and mitogen-activated kinase (MAPK) signaling proteins were determined by western blot analysis of UVB-irradiated HaCaT cells with or without prior SME treatment. In addition, the loss of mitochondrial membrane potential (Δ<I>ψ</I><SUB>m</SUB>) was detected using flow cytometry or confocal microscopy and the mitochondria membrane-permeate dye, JC-1. Apoptosis was assessed by quantifying DNA fragmentation and apoptotic body formation. Furthermore, cell viability was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.</P><P><I>Results</I>: SME absorbed electromagnetic radiation in the UVB range (280-320 nm) of the UV/visible light spectrum. SME also increased Bcl-2 and Mcl-1 expression in UVB-irradiated cells and decreased the Bax expression. Moreover, SME inhibited the UVB-induced disruption of mitochondrial membrane potential and prevented UVB-mediated increases in activated caspase-9 and caspase-3 (an apoptotic initiator and executor, respectively) levels. Notably, treatment with a pan-caspase inhibitor enhanced the anti-apoptotic effects of SME in UVB-irradiated cells. Finally, SME reduced the UVB-mediated phosphorylation of p38 MAPK and JNK, and prevented the UVB-mediated dephosphorylation of Erk1/2 and Akt.</P><P><I>Discussion and conclusion</I>: The present results indicate that SME safeguards HaCaT keratinocytes from UVB-mediated apoptosis by inhibiting a caspase-dependent signaling pathway.</P>

Chondracanthus tenellus (Harvey) hommersand extract protects the human keratinocyte cell line by blocking free radicals and UVB radiation-induced cell damage.

Piao, Mei Jing,Hyun, Yu Jae,Oh, Tae-Heon,Kang, Hee Kyoung,Yoo, Eun Sook,Koh, Young Sang,Lee, Nam Ho,Suh, In Soo,Hyun, Jin Won Springer 2012 In vitro cellular & developmental biology Animal Vol.48 No.10

<P>The aim of this study was to investigate the protective effects of the ethanol extract of the red algae Chondracanthus tenellus (Harvey) Hommersand (CTE) on cultured human keratinocyte cell line. The cellular protection conferred by CTE was evidenced by the ability of the extract to absorb ultraviolet B (UVB; 280-320 nm) and to scavenge the radical 1,1-diphenyl-2-picrylhydrazyl, as well as intracellular reactive oxygen species (ROS), induced by either hydrogen peroxide (H(2)O(2)) or UVB radiation. In addition, both superoxide anion generated by the xanthine/xanthine oxidase system and hydroxyl radical generated by the Fenton reaction (FeSO(4)?+?H(2)O(2)) were scavenged by CTE, as confirmed using electron spin resonance spectrometry. In the human keratinocyte cell line, CTE decreased the degree of injury resulting from UVB-induced oxidative stress to lipids, proteins, and DNA. CTE-treated cells also showed a reduction in UVB-induced apoptosis, as exemplified by fewer apoptotic bodies and less DNA fragmentation. Taken together, these results suggest that CTE confers protection on the human keratinocyte cell line against UVB-induced oxidative stress by absorbing UVB ray and scavenging ROS, thereby reducing injury to cellular constituents.</P>

Diphlorethohydroxycarmalol attenuated cell damage against UVB radiation <i>via</i> enhancing antioxidant effects and absorbing UVB ray in human HaCaT keratinocytes

Piao, Mei Jing,Kang, Kyoung Ah,Kim, Ki Cheon,Chae, Sungwook,Kim, Gi Ok,Shin, Taekyun,Kim, Hye Sun,Hyun, Jin Won Elsevier 2013 Environmental toxicology and pharmacology Vol.36 No.2

<P><B>Abstract</B></P> <P>Exposure of human skin to excessive ultraviolet B (UVB) radiation induces pathophysiological processes <I>via</I> the generation of reactive oxygen species (ROS) in skin cells, such as keratinocytes. This study investigated the ability of diphlorethohydroxycarmalol (DPHC) to protect human keratinocytes (HaCaT) against UVB-induced cell damage. DPHC restored cell viability that was reduced by UVB light. DPHC had an absorption maximum close to the UVB spectrum and decreased UVB-induced intracellular ROS levels, increased levels of reduced glutathione, activated superoxide dismutase and catalase. DPHC also decreased UVB-mediated damage to cellular components, including lipids, proteins, DNA, and attenuated UVB-induced apoptosis. These results suggest that DPHC safeguards human keratinocytes against UVB-induced cell damage by absorbing UVB ray, scavenging ROS and enhancing antioxidant system.</P> <P><B>Highlights</B></P> <P> <UL> <LI> DPHC had an absorption maximum close to the UVB spectrum. </LI> <LI> DPHC decreased UVB-induced intracellular ROS levels. </LI> <LI> DPHC increased reduced glutathione and antioxidant enzymes. </LI> <LI> DPHC decreased UVB-mediated damage to cellular components and attenuated apoptosis. </LI> <LI> DPHC protects human keratinocytes HaCaT against UVB-induced cell damage. </LI> </UL> </P>