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

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.

Combination Effect of Titrated Extract of Centella asiatica and Astaxanthin in a Mouse Model of Phthalic Anhydride-Induced Atopic Dermatitis

PURPOSE: In our previous study, we demonstrated that both titrated extract of Centella asiatica (TECA) and astaxanthin (AST) have anti-inflammatory effects in a 5% phthalic anhydride (PA) mouse model of atopic dermatitis (AD). The increasing prevalence of AD demands new therapeutic approaches for treating the disease. We investigated the therapeutic efficacy of the ointment form of TECA, AST and a TECA + AST combination in a mouse model of AD to see whether a combination of the reduced doses of 2 compounds could have a synergistic effect. METHODS: An AD-like lesion was induced by the topical application of 5% PA to the dorsal ear and back skin of an Hos:HR-1 mouse. After AD induction, TECA (0.5%), AST (0.5%) and the TECA (0.25%) + AST (0.25%) combination ointment (20 μg/cm2) were spread on the dorsum of the ear or back skin 3 times a week for 4 weeks. We evaluated dermatitis severity, histopathological changes and changes in protein expression by Western blotting for inducible nitric oxide synthase (iNOS), cyclocxygenase (COX)-2, and nuclear factor (NF)-κB activity. We also measured the concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and immunoglobulin E (IgE) in the blood of AD mice by enzyme-linked immunosorbent assay (ELISA). RESULTS: PA-induced skin morphological changes and ear thickness were significantly reduced by TECA, AST and TECA + AST treatments, but these inhibiting effects were more pronounced in the TECA + AST treatment. TECA, AST and the TECA+AST reatments inhibited the expression of iNOS and COX-2; NF-κB activity; and the release of TNF-α, IL-6 and IgE. However, the TECA+AST treatment showed additive or synergistic effects on AD. CONCLUSIONS: Our results demonstrate that the combination of TECA and AST could be a promising therapeutic agent for AD by inhibiting NF-κB signaling.

Anti-inflammatory effect of Centella asiatica phytosome in a mouse model of phthalic anhydride-induced atopic dermatitis.

BACKGROUND: Centella asiatica phytosome (CA phytosome) has potent antioxidant and anti-inflammatory properties. However, its anti-dermatitic effect has not yet been reported. PURPOSE: We investigated the effects of CA phytosome on inflammatory reponses by macrophages in an atopic dermatitis (AD) mouse model. STUDY DESIGN: The effects of CA phytosome on atopic dermatitis were examined by using phthalic anhydride (PA)-induced AD mouse model and RAW 264.7 murine macrophages. METHODS: An AD-like lesion was induced by a topical application of 5% phthalic anhydride (PA) to the dorsal skin or ear of HR-1 mice. After AD induction, 100 µl (20 µl/cm2) of 0.2% and 0.4% CA phytosome was spread on the dorsal skin and ear of the mice three times a week for four weeks. We evaluated histopathological changes and changes in protein expression by Western blotting for iNOS and COX-2; NF-κB activity was determined by EMSA. We also measured TNF-α, IL-1ß, and IgE concentration in the blood of AD mice by ELISA. RESULTS: Histological analysis showed that CA phytosome inhibited infiltration of inflammatory cells. CA phytosome treatment inhibited the expression of iNOS and COX-2, activity of NF-κB, and release of TNF-α, IL-1ß, and IgE. In addition, CA phytosome (5, 10, and 20 µg/ml) potently inhibited LPS (1 µg/ml)-induced NO production as well as iNOS and COX-2 expression in RAW 264.7 macrophage. Furthermore, CA phytosome inhibited LPS-induced DNA binding activities of NF-κB, and this was associated with the discontinuation of IκBα degradation and subsequent decreases in the translocation of p65 and p50 into the nucleus. CONCLUSION: From our data, CA phytosome application, which operates via NF-κB signaling inhibition, seems to be a promising AD treatment. Herein, we investigated the effects of Centella asiatica phytosome (CA phytosome) on inflammatory responses by macrophages in an atopic dermatitis (AD) mouse model. An AD-like lesion was induced by the topical application of 5% phthalic anhydride (PA) to the dorsal skin or ear of HR-1 mice. After AD induction, 100 µl (20 µl/cm2) of 0.2% and 0.4% CA phytosome was spread on the dorsal skin and ear of the mice three times a week for four weeks. We evaluated dermatitis severity, histopathological changes, and changes in protein expression by Western blotting for iNOS and COX-2; NF-κB activity was determined by gel electromobility shift assay (EMSA). We also measured TNF-α, IL-1ß, and IgE concentration in the blood of AD mice by enzyme-linked immunosorbent assay (ELISA). CA phytosome attenuated the development of PA-induced AD. Histological analysis showed that CA phytosome inhibited hyperkeratosis, proliferation of mast cells, and infiltration of inflammatory cells. Furthermore, CA phytosome treatment inhibited the expression of iNOS and COX-2, activity of NF-κB, and release of TNF-α, IL-1ß, and IgE. In addition, CA phytosome (5, 10, and 20 µg/ml) potently inhibited lipopolysaccharide (LPS) (1 µg/ml)-induced NO production as well as iNOS and COX-2 expression in RAW 264.7 macrophage cells. Furthermore, CA phytosome inhibited LPS-induced DNA binding activities of NF-κB, and this was associated with the discontinuation of IκBα degradation and subsequent decreases in the translocation of p65 and p50 into the nucleus. From our data, CA phytosome application, which operates via NF-κB signaling inhibition, seems to be a promising AD treatment.

Galangin (3,5,7-Trihydroxyflavone) Shields Human Keratinocytes from Ultraviolet B-Induced Oxidative Stress

Most skin damage caused by ultraviolet B (UVB) radiation is owing to the generation of reactive oxygen species. Phytochemicals can act as antioxidants against UVB-induced oxidative stress. This study investigated the protective effects of the flavone galangin against UVB-induced oxidative damage in human keratinocytes. Galangin efficiently scavenged free radicals and reduced UVB-induced damage to cellular macromolecules, such as DNA, lipids, and proteins. Furthermore, galangin rescued cells undergoing apoptosis induced by UVB radiation via recovering mitochondrial polarization and down-regulating apoptotic proteins. These results showed that galangin protects human keratinocytes against UVB radiation-induced cellular damage and apoptosis via its antioxidant effects.

Isorhamnetin Protects Human Keratinocytes against Ultraviolet B-Induced Cell Damage

Isorhamnetin (3-methylquercetin) is a flavonoid derived from the fruits of certain medicinal plants. This study investigated the photoprotective properties of isorhamnetin against cell damage and apoptosis resulting from excessive ultraviolet (UV) B exposure in human HaCaT keratinocytes. Isorhamnetin eliminated UVB-induced intracellular reactive oxygen species (ROS) and attenuated the oxidative modification of DNA, lipids, and proteins in response to UVB radiation. Moreover, isorhamnetin repressed UVB-facilitated programmed cell death in the keratinocytes, as evidenced by a reduction in apoptotic body formation, and nuclear fragmentation. Additionally, isorhamnetin suppressed the ability of UVB light to trigger mitochondrial dysfunction. Taken together, these results indicate that isorhamnetin has the potential to protect human keratinocytes against UVB-induced cell damage and death.

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

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.

The Polyphenol Chlorogenic Acid Attenuates UVB-mediated Oxidative Stress in Human HaCaT Keratinocytes

We investigated the protective effects of chlorogenic acid (CGA), a polyphenol compound, on oxidative damage induced by UVB exposure on human HaCaT cells. In a cell-free system, CGA scavenged 1,1-diphenyl-2-picrylhydrazyl radicals, superoxide anions, hydroxyl radicals, and intracellular reactive oxygen species (ROS) generated by hydrogen peroxide and ultraviolet B (UVB). Furthermore, CGA absorbed electromagnetic radiation in the UVB range (280-320 nm). UVB exposure resulted in damage to cellular DNA, as demonstrated in a comet assay; pre-treatment of cells with CGA prior to UVB irradiation prevented DNA damage and increased cell viability. Furthermore, CGA pre-treatment prevented or ameliorated apoptosis-related changes in UVB-exposed cells, including the formation of apoptotic bodies, disruption of mitochondrial membrane potential, and alterations in the levels of the apoptosis-related proteins Bcl-2, Bax, and caspase-3. Our findings suggest that CGA protects cells from oxidative stress induced by UVB radiation.

Fucodiphlorethol G Purified from Ecklonia cava Suppresses Ultraviolet B Radiation-Induced Oxidative Stress and Cellular Damage

Fucodiphlorethol G (6'-[2,4-dihydroxy-6-(2,4,6-trihydroxyphenoxy)phenoxy]biphenyl-2,2',4,4',6-pentol) is a compound purified from Ecklonia cava, a brown alga that is widely distributed offshore of Jeju Island. This study investigated the protective effects of fucodiphlorethol G against oxidative damage-mediated apoptosis induced by ultraviolet B (UVB) irradiation. Fucodiphlorethol G attenuated the generation of 2, 2-diphenyl-1-picrylhydrazyl radicals and intracellular reactive oxygen species in response to UVB irradiation. Fucodiphlorethol G suppressed the inhibition of human keratinocyte growth by UVB irradiation. Additionally, the wavelength of light absorbed by fucodiphlorethol G was close to the UVB spectrum. Fucodiphlorethol G reduced UVB radiation-induced 8-isoprostane generation and DNA fragmentation in human keratinocytes. Moreover, fucodiphlorethol G reduced UVB radiation-induced loss of mitochondrial membrane potential, generation of apoptotic cells, and active caspase-9 expression. Taken together, fucodiphlorethol G protected human keratinocytes against UVB radiation-induced cell damage and apoptosis by absorbing UVB radiation and scavenging reactive oxygen species.

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

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 flavonoid fisetin (3,7,3',4'-tetrahydroxyflavone) included the radioprotection of cells exposed to gamma-irradiation. Fisetin reduced the levels of intracellular reactive oxygen species generated by gamma-irradiation and thereby protected cells against gamma-irradiation-induced membrane lipid peroxidation, DNA damage, and protein carbonylation. In addition, fisetin maintained the viability of irradiated cells by partially inhibiting gamma-irradiation-induced apoptosis and restoring mitochondrial membrane potential. These effects suggest that the cellular protective effects of fisetin against gamma-irradiation are mainly due to its inhibition of reactive oxygen species generation.

6'-O-Galloylpaeoniflorin Protects Human Keratinocytes Against Oxidative Stress-Induced Cell Damage

6'-O-galloylpaeoniflorin (GPF) is a galloylated derivate of paeoniflorin and a key chemical constituent of the peony root, a perennial flowering plant that is widely used as an herbal medicine in East Asia. This study is the first investigation of the cytoprotective effects of GPF against hydrogen peroxide (H2O2)-induced cell injury and death in human HaCaT keratinocytes. GPF demonstrated a significant scavenging capacity against the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, H2O2-generated intracellular reactive oxygen species (ROS), the superoxide anion radical (O2-), and the hydroxyl radical (*OH). GPF also safeguarded HaCaT keratinocytes against H2O2-provoked apoptotic cell death and attenuated oxidative macromolecular damage to DNA, lipids, and proteins. The compound exerted its cytoprotective actions in keratinocytes at least in part by decreasing the number of DNA strand breaks, the levels of 8-isoprostane (a stable end-product of lipid peroxidation), and the formation of carbonylated protein species. Taken together, these results indicate that GPF may be developed as a cytoprotector against ROS-mediated oxidative stress.

Protective effect of the methanol extract of Polyopes lancifolia (Harvey) kawaguchi et wang against ionizing radiation-induced mouse gastrointestinal injury / 대한수의학회지

The radioprotective efficacy of a methanol extract of the red algae Polyopes lancifolia (Harvey) kawaguchi et wang (mPL) was evaluated in mice subjected to total-body gamma irradiation. mPL protection against radiation-induced oxidative stress was examined by histological evaluation of intestinal crypt-cell survival and liver activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). mPL (100 mg/kg body weight) administered intraperitoneally at 24 h and 1 h prior to irradiation protected jejunal crypt cells from radiation-induced apoptosis (p < 0.01). The pretreatment of mPL attenuated a radiation-induced decrease in villous height (p < 0.05), and improved jejunal crypt survival (p < 0.05). The dose reduction factor was 1.14 at 3.5 days after irradiation. Treatment with mPL prior to irradiation resulted in significantly higher (p < 0.01) levels of SOD and CAT activities, compared to those levels of irradiated control mice with vehicle treatment. These results suggest that mPL is a useful radioprotective agent capable of defending intestinal progenitor cells against total-body irradiation, at least in part through mPL antioxidative activity.