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Photoprotective effects of phloretin sulfonate against UVB-induced damage in skin cell model and human volunteers
Abstracts / Journal of Dermatological Science 84 (2016) e1–e88
acid residues of SC proteins resulting in the formation of carbonylated proteins. Carbonylated proteins in the SC are more frequently detected in the outermost layer than in the inner layer. We found increased levels of carbonylated proteins in SC transferred on glass slides after UV irradiation in an in vitro study. In addition, it has been reported that ROS levels increased by UVA irradiation decline with the repeated tape-stripping of the SC. These facts suggest that the SC when UV irradiated might play a role as a generation source of ROS. In this study, we examined the increase of chemiluminescence intensity of MCLA to detect O2 − and 1 O2 from UV-irradiated SC. Furthermore, the type of ROS produced is suggested to be·O2 − since the addition of SOD to the system resulted in an attenuation of the chemiluminescence. The results of this study demonstrate the involvement of carbonylated proteins in the generation of ROS in the SC. http://dx.doi.org/10.1016/j.jdermsci.2016.08.242 P12-07 Inflammatory actions of near-infrared radiation by increasing MMP-1 and IL-8 production in human dermal fibroblasts Hiroko Sugimoto 1,∗ , Noriko Akimoto 1 , Shiho Tanaka 2 , Mana Hirayama 3 , Yuko Takenaka 3 , Yuichiro Tsunemi 3 , Makoto Kawashima 3 , Takashi Sato 1 1
Department of Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan 2 Aoyama Institute of Women’s Medicine, Tokyo Women’s Medical University, Tokyo, Japan 3 Department of Dermatology, Tokyo Women’s Medical University, Tokyo, Japan Objective: Near-infrared radiation (NIR), which is a composition of sunlight with a 800–2000 nm wavelength, is associated with skin aging such as wrinkles and slacks. On the other hand, NIR irradiation is effective for the improvement of the skin photo damage in humans. In the present study, as a new NIR irradiation apparatus that controls wave length and irradiation dose has been developed, we examined the effect of NIR on cellular functions of human dermal fibroblasts (HDF) using the apparatus. Methods: The NIR irradiation apparatus has a xenon flash lamp, of which light is white near that of sunlight (900–1700 nm). HDF were irradiated for 3.5–7 min with NIR (190 mW/cm2 ) in the presence or absence of a filter intercepting 900–1000 nm wavelength, and then maintained for 24 h. The production and mRNA expression of matrix metalloproteinase 1 (MMP-1), MMP-2, MMP-3, and interleukin 8 (IL-8) were monitored by Western blotting, ELISA, gelatin zymography, and real-time PCR. Results: NIR irradiation was found to transcriptionally augment the production of MMP-1 and IL-8 in HDF. However, the gene expression and production of MMPs-2 and -3 in HDF were not influenced under the same conditions. On the other hand, NIR irradiation using a filter (1000–1700 nm) no longer increased the production and gene expression of MMP-1 and IL-8 in HDF, as well as those of MMPs-2 and -3. Conclusion: These results suggest novel evidence that a narrow wavelength range of NIR (900–1000 nm) exhibits inflammatory actions due to the transcriptional augmentation of MMP-1 and IL-8 production in HDF. Furthermore, NIR irradiation is likely to selectively up-regulate the expression of MMP-1 rather than that of MMPs-2 and -3 in HDF. http://dx.doi.org/10.1016/j.jdermsci.2016.08.243
e79
P12-08 Photoprotective effects of phloretin sulfonate against UVB-induced damage in skin cell model and human volunteers Seoung Woo Shin ∗ , Hyunwoo Kum, Minkyung Kim, Eunsun Jung, Deokhoon Park BioSpectrum, Inc., Seongnam, Republic of Korea The phenolic compound phloretin is a prominent member of the chemical class of dihydrochalcones. Phloretin is specifically found in apple and apple juice and known for its biological properties. In particular we were attracted by potential dermocosmetic applications. However, practical limitations of phloretin do exist due to its poor water-solubility. To increase its watersolubility, phloretin was sulfonated with sulfuric acid (98 wt.%) and mixed with saturated salt water to produce phloretin sodium sulfonate. Here we report the photoprotective effect of phloretin sulfonate, a new semi-synthetic derivative of phloretin. Results showed that phloretin sulfonate can attenulate cyclobutane pyrimidine dimers (CPDs) formation, glutathione (GSH) depletion and apoptosis induced by UVB. The photoprotective effect of phloretin sulfonate is tightly correlated to the enhancement of NER genes expression. Furthermore, phloretin sulfonate had inhibitory effects on the UVB-induced release of the inflammatory mediators such as IL-6 and prostaglandin-E2. We also confirmed the safety and clinical efficacy of phloretin sulfonate on human skin. Overall, results demonstrated significant benefits of phloretin sulfonate on the protection of keratinocyte against UVB-induced injuries and suggested its potential use in skin photoprotection. http://dx.doi.org/10.1016/j.jdermsci.2016.08.244 P12-09 Microarray analysis in the keratinocyte and melanocyte exposed to narrow-band UVB and broad-band UVB Taro Masaki 1,∗ , Seiji Takeuchi 1 , Toshiro Matsuda 2 , Chikako Nishigori 1 1 Division of Dermatology, Clinical Molecular Medicine, Faculty of Medicine, Kobe University Graduate School of Medicine, Kobe, Japan 2 Atomic Energy Research Institute, Kinki University, Osaka, Japan
Phototherapy using narrow-band UVB (NB-UVB) has been used for various skin diseases such as psoriasis, vitiligo and atopic dermatitis. While it has a beneficial effect in treating these patients, in animal model of UV carcinogenesis, NB-UVB develops much more malignant skin tumors than broad-band UVB (BB-UVB) and it develops skin tumors more rapidly than BB-UVB. To investigate the effect of NB-UVB and BB-UVB, we analyzed gene expression profile using microarray of human melanocytes (MCs) and keratinocytes (KCs) after NB-UVB and BB-UVB exposure. Total RNAs from human MCs and KCs were isolated at 3hr after NB-UVB exposure at 3 kJ/m2 or BB-UVB exposure at 300 J/m2 and the data were analyzed with software Genespring 12.5. Genes with a fold difference of >2.0 or <0.5 were considered significant. All transcripts data were compared to the expression of the counterpart of unirradiated samples. The results showed that the number of gene variations after BB-UVB exposure was much more than those after NB-UVB exposure in both MCs and KCs, and those may reflect the difference in the efficacy and side effect of the NB-UVB and BB-UVB. Moreover, there was a considerable difference between the pathways of KC and MC about
acid residues of SC proteins resulting in the formation of carbonylated proteins. Carbonylated proteins in the SC are more frequently detected in the outermost layer than in the inner layer. We found increased levels of carbonylated proteins in SC transferred on glass slides after UV irradiation in an in vitro study. In addition, it has been reported that ROS levels increased by UVA irradiation decline with the repeated tape-stripping of the SC. These facts suggest that the SC when UV irradiated might play a role as a generation source of ROS. In this study, we examined the increase of chemiluminescence intensity of MCLA to detect O2 − and 1 O2 from UV-irradiated SC. Furthermore, the type of ROS produced is suggested to be·O2 − since the addition of SOD to the system resulted in an attenuation of the chemiluminescence. The results of this study demonstrate the involvement of carbonylated proteins in the generation of ROS in the SC. http://dx.doi.org/10.1016/j.jdermsci.2016.08.242 P12-07 Inflammatory actions of near-infrared radiation by increasing MMP-1 and IL-8 production in human dermal fibroblasts Hiroko Sugimoto 1,∗ , Noriko Akimoto 1 , Shiho Tanaka 2 , Mana Hirayama 3 , Yuko Takenaka 3 , Yuichiro Tsunemi 3 , Makoto Kawashima 3 , Takashi Sato 1 1
Department of Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan 2 Aoyama Institute of Women’s Medicine, Tokyo Women’s Medical University, Tokyo, Japan 3 Department of Dermatology, Tokyo Women’s Medical University, Tokyo, Japan Objective: Near-infrared radiation (NIR), which is a composition of sunlight with a 800–2000 nm wavelength, is associated with skin aging such as wrinkles and slacks. On the other hand, NIR irradiation is effective for the improvement of the skin photo damage in humans. In the present study, as a new NIR irradiation apparatus that controls wave length and irradiation dose has been developed, we examined the effect of NIR on cellular functions of human dermal fibroblasts (HDF) using the apparatus. Methods: The NIR irradiation apparatus has a xenon flash lamp, of which light is white near that of sunlight (900–1700 nm). HDF were irradiated for 3.5–7 min with NIR (190 mW/cm2 ) in the presence or absence of a filter intercepting 900–1000 nm wavelength, and then maintained for 24 h. The production and mRNA expression of matrix metalloproteinase 1 (MMP-1), MMP-2, MMP-3, and interleukin 8 (IL-8) were monitored by Western blotting, ELISA, gelatin zymography, and real-time PCR. Results: NIR irradiation was found to transcriptionally augment the production of MMP-1 and IL-8 in HDF. However, the gene expression and production of MMPs-2 and -3 in HDF were not influenced under the same conditions. On the other hand, NIR irradiation using a filter (1000–1700 nm) no longer increased the production and gene expression of MMP-1 and IL-8 in HDF, as well as those of MMPs-2 and -3. Conclusion: These results suggest novel evidence that a narrow wavelength range of NIR (900–1000 nm) exhibits inflammatory actions due to the transcriptional augmentation of MMP-1 and IL-8 production in HDF. Furthermore, NIR irradiation is likely to selectively up-regulate the expression of MMP-1 rather than that of MMPs-2 and -3 in HDF. http://dx.doi.org/10.1016/j.jdermsci.2016.08.243
e79
P12-08 Photoprotective effects of phloretin sulfonate against UVB-induced damage in skin cell model and human volunteers Seoung Woo Shin ∗ , Hyunwoo Kum, Minkyung Kim, Eunsun Jung, Deokhoon Park BioSpectrum, Inc., Seongnam, Republic of Korea The phenolic compound phloretin is a prominent member of the chemical class of dihydrochalcones. Phloretin is specifically found in apple and apple juice and known for its biological properties. In particular we were attracted by potential dermocosmetic applications. However, practical limitations of phloretin do exist due to its poor water-solubility. To increase its watersolubility, phloretin was sulfonated with sulfuric acid (98 wt.%) and mixed with saturated salt water to produce phloretin sodium sulfonate. Here we report the photoprotective effect of phloretin sulfonate, a new semi-synthetic derivative of phloretin. Results showed that phloretin sulfonate can attenulate cyclobutane pyrimidine dimers (CPDs) formation, glutathione (GSH) depletion and apoptosis induced by UVB. The photoprotective effect of phloretin sulfonate is tightly correlated to the enhancement of NER genes expression. Furthermore, phloretin sulfonate had inhibitory effects on the UVB-induced release of the inflammatory mediators such as IL-6 and prostaglandin-E2. We also confirmed the safety and clinical efficacy of phloretin sulfonate on human skin. Overall, results demonstrated significant benefits of phloretin sulfonate on the protection of keratinocyte against UVB-induced injuries and suggested its potential use in skin photoprotection. http://dx.doi.org/10.1016/j.jdermsci.2016.08.244 P12-09 Microarray analysis in the keratinocyte and melanocyte exposed to narrow-band UVB and broad-band UVB Taro Masaki 1,∗ , Seiji Takeuchi 1 , Toshiro Matsuda 2 , Chikako Nishigori 1 1 Division of Dermatology, Clinical Molecular Medicine, Faculty of Medicine, Kobe University Graduate School of Medicine, Kobe, Japan 2 Atomic Energy Research Institute, Kinki University, Osaka, Japan
Phototherapy using narrow-band UVB (NB-UVB) has been used for various skin diseases such as psoriasis, vitiligo and atopic dermatitis. While it has a beneficial effect in treating these patients, in animal model of UV carcinogenesis, NB-UVB develops much more malignant skin tumors than broad-band UVB (BB-UVB) and it develops skin tumors more rapidly than BB-UVB. To investigate the effect of NB-UVB and BB-UVB, we analyzed gene expression profile using microarray of human melanocytes (MCs) and keratinocytes (KCs) after NB-UVB and BB-UVB exposure. Total RNAs from human MCs and KCs were isolated at 3hr after NB-UVB exposure at 3 kJ/m2 or BB-UVB exposure at 300 J/m2 and the data were analyzed with software Genespring 12.5. Genes with a fold difference of >2.0 or <0.5 were considered significant. All transcripts data were compared to the expression of the counterpart of unirradiated samples. The results showed that the number of gene variations after BB-UVB exposure was much more than those after NB-UVB exposure in both MCs and KCs, and those may reflect the difference in the efficacy and side effect of the NB-UVB and BB-UVB. Moreover, there was a considerable difference between the pathways of KC and MC about