Rosmarinic Acid Inhibits Ultraviolet B-Mediated Oxidative Damage via the AKT/ERK-NRF2-GSH Pathway In Vitro and In Vivo

( Mei Jing Piao ),( Pattage Madushan Dilhara Jayatissa Fernando ),( Kyoung Ah Kang ),( Pincha Devage Sameera Madushan Fernando ),( Herath Mudiyanselage Udari Lakmini Herath ),( Young Ree Kim ),( Jin W 한국응용약물학회 2024 Biomolecules & Therapeutics(구 응용약물학회지) Vol.32 No.1

Rosmarinic acid (RA) is a phenolic ester that protects human keratinocytes against oxidative damage induced by ultraviolet B (UVB) exposure, however, the mechanisms underlying its effects remain unclear. This study aimed to elucidate the cell signaling mechanisms that regulate the antioxidant activity of RA and confirm its cyto-protective role. To explore the signaling mechanisms, we used the human keratinocyte cell line HaCaT and SKH1 hairless mouse skin. RA enhanced glutamate-cysteine ligase catalytic subunit (GCLC) and glutathione synthetase (GSS) expression in HaCaT cells in a dose- and time-dependent manner. Moreover, RA induced nuclear factor erythroid-2-related factor 2 (NRF2) nuclear translocation and activated the signaling kinases protein kinase B (AKT) and extracellular signal-regulated kinase (ERK). Treatment with the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002, the ERK inhibitor U0126, and small interfering RNA (siRNA) gene silencing suppressed RA-enhanced GCLC, GSS, and NRF2 expression, respectively. Cell viability tests showed that RA significantly prevented UVB-induced cell viability decrease, whereas the glutathione (GSH) inhibitors buthionine sulfoximine, LY294002, and U0126 significantly reduced this effect. Moreover, RA protected against DNA damage and protein carbonylation, lipid peroxidation, and apoptosis caused by UVB-induced oxidative stress in a concentration-dependent manner in SKH1 hairless mouse skin tissues. These results suggest that RA protects against UVB-induced oxidative damage by activating AKT and ERK signaling to regulate NRF2 signaling and enhance GSH biosynthesis. Thus, RA treatment may be a promising approach to protect the skin from UVB-induced oxidative damage.

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>

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>

Phloroglucinol Attenuates Ultraviolet B Radiation‐Induced Matrix Metalloproteinase‐1 Production in Human Keratinocytes <i>via</i> Inhibitory Actions against Mitogen‐Activated Protein Kinases and Activator Protein‐1

Piao, Mei Jing,Zhang, Rui,Lee, Nam Ho,Hyun, Jin Won Blackwell Publishing Ltd 2012 Photochemistry and photobiology Vol.88 No.2

<P><B>Abstract</B></P><P>Excessive amounts of reactive oxygen species (ROS) induced by ultraviolet (UV) radiation cause skin aging <I>via</I> basement membrane/extracellular matrix degradation resulting from the action of matrix metalloproteinases (MMPs). Recently, phloroglucinol (1,3,5‐trihydroxybenzene) was demonstrated to attenuate the cell damage induced by oxidative stress by quenching ROS and stimulating antioxidant systems. In the current study, the effect of phloroglucinol on UVB‐induced photoaging was investigated in human HaCaT keratinocytes. Phloroglucinol significantly inhibited the UVB‐induced (1) upregulation of MMP‐1 mRNA, protein and activity; (2) augmentation of intracellular Ca<SUP>2+</SUP> levels; (3) phosphorylation of mitogen‐activated protein kinases (MAPKs); (4) expression of c‐Fos and phospho c‐Jun; and (5) enhancement of activator protein‐1 (AP‐1) binding to the MMP‐1 promoter. In addition, the knockdown of MAPKs significantly inhibited UVB‐induced MMP‐1 expression. The results of this study suggest that phloroglucinol may be useful as a photoprotective compound for the skin.</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>

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>

Photo-protective Effect of Polysiphonia morrowii Harvey against Ultraviolet B Radiation-induced Keratinocyte Damage

Piao, Mei Jing,Kang, Hee-Kyoung,Yoo, Eun-Sook,Koh, Young-Sang,Kim, Dong-Sam,Lee, Nam-Ho,Hyun, Jin-Won The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.2

Photo-protective properties of Polysiphonia morrowii ethanol extract (PME) against ultraviolet B (UVB) radiation-induced cellular damage were investigated in human HaCaT keratinocytes. PME exhibited scavenging activity against 1,1-diphenyl-2-picrylhydrazyl radical and intracellular reactive oxygen species induced by either hydrogen peroxide ($H_2O_2$) or UVB radiation. In addition, PME scavenged superoxide anion generated by xanthine/xanthine oxidase system and hydroxyl radical generated by the Fenton reaction ($FeSO_4+H_2O_2$), both of which were detected using electron spin resonance spectrometry. Moreover, PME, which scavenges $H_2O_2$, was shown to contain the antioxidant compound 3-bromo-4,5-dihydroxylbenzaldehyde, possibly accounting for scavenging actions. PME also restored the levels of catalase and superoxide dismutase protein expression and activity that were decreased by UVB radiation. PME reduced UVB-induced apoptosis, as shown by decreased apoptotic bodies and DNA fragmentation. These results indicate that PME protects human keratinocytes against oxidative stress induced by UVB radiation through the enhancement of antioxidant systems.

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>

Eckol inhibits ultraviolet B-induced cell damage in human keratinocytes via a decrease in oxidative stress.

Piao, Mei Jing,Lee, Nam Ho,Chae, Sungwook,Hyun, Jin Won Pharmaceutical Society of Japan 2012 BIOLOGICAL & PHARMACEUTICAL BULLETIN Vol.35 No.6

<P>In previous reports, the antioxidant effects of eckol were shown to protect cells against hydrogen peroxide- and gamma ray-induced oxidative stress. In this study, the role of eckol in protecting human skin keratinocytes (HaCaT) against UVB-induced oxidative cell damage was investigated. Also, triphlorethol-A, one of the chemical components in Ecklonia cava, and quercetin a well known antioxidant, were compared with eckol in terms of antioxidant activity based on chemical structure. Eckol decreased UVB-induced intracellular reactive oxygen species (ROS), decreased injury to cellular components resulting from UVB-induced oxidative stress, and restored cell viability. In addition, eckol reduced UVB-induced apoptosis by inhibiting the disruption of mitochondrial membranes. These results suggest that eckol protects human keratinocytes against UVB-induced oxidative stress by scavenging ROS, thereby lessening injury to cellular components.</P>

Hyperoside Protects Cells against Gamma Ray Radiation-Induced Apoptosis in Hamster Lung Fibroblast

Mei Jing Piao,현진원,김기천,조석주,채성욱,강삼식 한국생약학회 2013 Natural Product Sciences Vol.19 No.2

Ionizing radiation, including that evoked by gamma (g)-rays, induces oxidative stress through the generation of reactive oxygen species, resulting in apoptosis, or programmed cell death. This study aimed to elucidate the radioprotective effects of hyperoside (quercetin-3-O-galactoside) against g-ray radiation-induced apoptosis in Chinese hamster lung fibroblasts, V79-4 and demonstrated that the compound reduced levels of intracellular reactive oxygen species in g-ray-irradiated cells. Hyperoside also protected irradiated cells against DNA damage (evidenced by pronounced DNA tails and elevated phospho-histone H2AX and 8-oxoguanine content) and membrane lipid peroxidation. Furthermore, hyperoside prevented the g-ray-provoked reduction in cell viability via the inhibition of apoptosis through the increased levels of Bcl-2, the decreased levels of Bax and cytosolic cytochrome c, and the decrease of the active caspase 9 and caspase 3 expression. Taken together, these results suggest that hyperoside defend cells against g-ray radiation-induced apoptosis by inhibiting oxidative stress.