Synergistic Cytoprotective Effects of Rutin and Ascorbic Acid on the Proteomic Profile of 3D-Cultured Keratinocytes Exposed to UVA or UVB Radiation

The combination of ascorbic acid and rutin, often used in oral preparations, due to antioxidant and anti-inflammatory properties, can be used to protect skin cells against the effects of UV radiation from sunlight. Therefore, the aim of this study was to investigate the synergistic effect of rutin and ascorbic acid on the proteomic profile of UVA and UVB irradiated keratinocytes cultured in a three-dimensional (3D) system. Results showed that the combination of rutin and ascorbic acid protects skin cells against UV-induced changes. In particular, alterations were observed in the expression of proteins involved in the antioxidant response, DNA repairing, inflammation, apoptosis, and protein biosynthesis. The combination of rutin and ascorbic acid also showed a stronger cytoprotective effect than when using either compound alone. Significant differences were visible between rutin and ascorbic acid single treatments in the case of protein carboxymethylation/carboxyethylation. Ascorbic acid prevented UV or rutin-induced protein modifications. Therefore, the synergistic effect of rutin and ascorbic acid creates a potentially effective protective system against skin damages caused by UVA and UVB radiation.

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Effects of Natural Antioxidants on Phospholipid and Ceramide Profiles of 3D-Cultured Skin Fibroblasts Exposed to UVA or UVB Radiation

Antioxidants ◽

10.3390/antiox10040578 ◽

2021 ◽

Vol 10 (4) ◽

pp. 578

Author(s):

Agnieszka Gęgotek ◽

Wojciech Łuczaj ◽

Elżbieta Skrzydlewska

Keyword(s):

Ascorbic Acid ◽

High Performance ◽

Three Dimensional ◽

Natural Antioxidants ◽

Phospholipid Metabolism ◽

Culture Conditions ◽

Membrane Properties ◽

Dimensional System ◽

Uvb Radiation ◽

Liquid Chromatograph

Ultraviolet (UV) radiation is one of the primary factors responsible for disturbances in human skin cells phospholipid metabolism. Natural compounds that are commonly used to protect skin, due to their lipophilic or hydrophilic nature, show only a narrow range of cytoprotective activity, which prompts research on their combined application. Therefore, the aim of this study was to examine the effect of ascorbic acid and rutin on the phospholipid and ceramide profiles in UV-irradiated fibroblasts cultured in a three-dimensional system that approximates the culture conditions to the dermis. An ultra-high-performance liquid chromatograph coupled with a quadrupole time-of-flight mass spectrometer was used for phospholipid and ceramide profiling. As a result of UVA and UVB cells irradiation, upregulation of phosphatidylcholines, ceramides, and downregulation of sphingomyelins were observed, while treatment with ascorbic acid and rutin of UVA/UVB-irradiated fibroblast promoted these changes to provide cells a stronger response to stress. Moreover, an upregulation of phosphatidylserines in cells exposed to UVB and treated with both antioxidants suggests the stimulation of UV-damaged cells apoptosis. Our findings provide new insight into action of rutin and ascorbic acid on regulation of phospholipid metabolism, which improves dermis fibroblast membrane properties.

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Cytoprotective Effect of Ascorbic Acid and Rutin against Oxidative Changes in the Proteome of Skin Fibroblasts Cultured in a Three-Dimensional System

Nutrients ◽

10.3390/nu12041074 ◽

2020 ◽

Vol 12 (4) ◽

pp. 1074 ◽

Cited By ~ 5

Author(s):

Agnieszka Gęgotek ◽

Iwona Jarocka-Karpowicz ◽

Elżbieta Skrzydlewska

Keyword(s):

Ascorbic Acid ◽

Uv Radiation ◽

Three Dimensional ◽

Skin Fibroblasts ◽

Combined Effect ◽

Dimensional System ◽

Protective Effects ◽

Skin Damage ◽

Skin Cells ◽

Extracellular Signaling

The combination of ascorbic acid and rutin, commonly used in oral preparations for their antioxidant and anti-inflammatory properties, can also be used to protect skin cells from the effects of UV radiation in sunlight. Here, we tested the potential protective effect of ascorbic acid and rutin used together in UVB-irradiated human skin fibroblasts, and assessed the proteomic profile of these cells, grown in a three-dimensional (3D) system. Proteomic findings revealed a combined effect of ascorbic acid and rutin in UV-irradiated fibroblasts against overexpression of pro-inflammatory signaling proteins and DNA reorganization/expression. These effects were not observed when cells were treated with either compounds alone. The antioxidant effects of ascorbic acid and rutin also prevented protein modifications by lipid peroxidation products. Further, ascorbic acid stimulated rutin-protein adduct formation, which supports intra/extracellular signaling and the Nrf2/ARE antioxidant pathway, contributing to the protective effects against UV-induced oxidative stress. The combined effect of ascorbic acid and rutin suggests that this combination of compounds is potentially effective against skin damage caused by UV radiation.

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The Differences in the Proteome Profile of Cannabidiol-Treated Skin Fibroblasts following UVA or UVB Irradiation in 2D and 3D Cell Cultures

Cells ◽

10.3390/cells8090995 ◽

2019 ◽

Vol 8 (9) ◽

pp. 995 ◽

Cited By ~ 8

Author(s):

Agnieszka Gęgotek ◽

Sinemyiz Atalay ◽

Pedro Domingues ◽

Elżbieta Skrzydlewska

Keyword(s):

Cultured Cells ◽

Uv Light ◽

Cell Metabolism ◽

Antioxidant Response ◽

Skin Fibroblasts ◽

Uvb Radiation ◽

Cultured Fibroblasts ◽

Skin Cells ◽

Skin Cell ◽

2D And 3D

Cannabidiol (CBD), as the only phytocannabinoid that has no psychoactive effect, has both antioxidant and anti-inflammatory effects, and thus might be suggested as a cytoprotective compound against UV-induced metabolic changes in skin cells. Therefore, the aim of this study was to investigate the level of protective CBD activity by evaluating the proteomic profile of 2D and 3D cultured skin fibroblasts models following exposure to UVA and UVB radiation. The CBD cytoprotective effect against UV-induced damage in 2D and 3D cultured fibroblasts were different. The main alterations focus on the range of cell reaction and involved different proteins associated with various molecular functions. In the 2D cultured cells, following UV radiation, the major changes were associated with proteins involved in antioxidant response and inflammation, while, in the 3D cultured fibroblasts, CBD action against UV induced changes were mainly associated with the activation of signalling pathways. Therefore, the knowledge of the CBD action in a multilayer skin cells model allowed for the prediction of changes in cell-cell interactions and skin cell metabolism. Knowledge about the lower protective effect of CBD in 3D cultured fibroblasts should be taken into account during the design of UV light protection.

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Conformation of Ribosomes from Escherichia Coli

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100051062 ◽

1974 ◽

Vol 32 ◽

pp. 210-211

Author(s):

M. Boublik ◽

W. Hellmann ◽

F. Jenkins

Keyword(s):

Binding Sites ◽

Ribosomal Proteins ◽

Fluorescent Dyes ◽

Protein Biosynthesis ◽

Three Dimensional ◽

Spatial Arrangement ◽

Dimensional Structure ◽

Complete Understanding ◽

And Function

The present knowledge of the three-dimensional structure of ribosomes is far too limited to enable a complete understanding of the various roles which ribosomes play in protein biosynthesis. The spatial arrangement of proteins and ribonuclec acids in ribosomes can be analysed in many ways. Determination of binding sites for individual proteins on ribonuclec acid and locations of the mutual positions of proteins on the ribosome using labeling with fluorescent dyes, cross-linking reagents, neutron-diffraction or antibodies against ribosomal proteins seem to be most successful approaches. Structure and function of ribosomes can be correlated be depleting the complete ribosomes of some proteins to the functionally inactive core and by subsequent partial reconstitution in order to regain active ribosomal particles.

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Cannabidiol effect on the proteomic profile of keratinocytes isolated from rat skin exposed to UVA and UVB radiation

Free Radical Biology and Medicine ◽

10.1016/j.freeradbiomed.2020.12.369 ◽

2021 ◽

Vol 165 ◽

pp. 40

Author(s):

Sinemyiz Atalay ◽

Agnieszka Gęgotek ◽

Pedro Domingues ◽

Elżbieta Skrzydlewska

Keyword(s):

Proteomic Profile ◽

Uvb Radiation ◽

Rat Skin

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Ascorbic acid and rutin cooperation in protecting of the proteome of UV irradiated fibroblasts cultured in a three-dimensional system

Free Radical Biology and Medicine ◽

10.1016/j.freeradbiomed.2020.12.368 ◽

2021 ◽

Vol 165 ◽

pp. 40

Author(s):

Agnieszka Gęgotek ◽

Pedro Domingues ◽

Elżbieta Skrzydlewska

Keyword(s):

Ascorbic Acid ◽

Three Dimensional ◽

Dimensional System ◽

Three Dimensional System

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Exogenous Antioxidants Impact on UV-Induced Changes in Membrane Phospholipids and the Effectiveness of the Endocannabinoid System in Human Skin Cells

Antioxidants ◽

10.3390/antiox10081260 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1260

Author(s):

Agnieszka Gęgotek ◽

Anna Jastrząb ◽

Marta Dobrzyńska ◽

Michał Biernacki ◽

Elżbieta Skrzydlewska

Keyword(s):

Ascorbic Acid ◽

Cell Viability ◽

Human Skin ◽

Phospholipid Fatty Acid ◽

Antioxidant Properties ◽

Endocannabinoid System ◽

Membrane Phospholipids ◽

Skin Cells ◽

Human Skin Cells ◽

Induced Changes

Natural antioxidants effectively counteract changes caused by UV radiation in human skin cells. However, their action is limited due to their lipo/hydrophilicity. Therefore, the aim of this study was to analyze the mutual protective action of hydrophilic ascorbic acid and partially lipophilic rutin against UVA/UVB-induced changes in membranes phospholipid and endocannabinoid system in keratinocytes and fibroblasts. Obtained results clearly showed that, despite the stronger antioxidant properties of ascorbic acid, the lipid membranes were more effectively protected against UV-induced oxidation by rutin, including changes in phospholipid fatty acid levels, prevention against reactive aldehydes formation and endocannabinoids degradation. Ascorbic acid more strongly prevented UV-induced endocannabinoid receptors expression in fibroblasts, especially CB1. However, the combined action of used antioxidants resulted in the greatest cytoprotective effect, which was evident in the inflammatory marker TNFα down-regulation and increased cell viability following cell irradiation. The applied mixture of antioxidants showed a stronger protective in relation to membrane phospholipids in keratinocytes and in the endocannabinoid system in fibroblasts. In conclusion, it can be suggested that combined antioxidant capacities of ascorbic acid and rutin protects against lipid peroxidation but also decreases the UV-induced inflammation by direct interaction with the endocannabinoid system, thus increasing skin cell viability.

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Marine Compound 3-bromo-4,5-dihydroxybenzaldehyde Protects Skin Cells against Oxidative Damage via the Nrf2/HO-1 Pathway

Marine Drugs ◽

10.3390/md17040234 ◽

2019 ◽

Vol 17 (4) ◽

pp. 234 ◽

Cited By ~ 3

Author(s):

Yea Seong Ryu ◽

Pincha Devage Sameera Madushan Fernando ◽

Kyoung Ah Kang ◽

Mei Jing Piao ◽

Ao Xuan Zhen ◽

...

Keyword(s):

Oxidative Damage ◽

Antioxidant Response ◽

Signaling Cascades ◽

Heme Oxygenase 1 ◽

Related Factor ◽

Skin Cells ◽

Reverse Transcription Pcr ◽

Extracellular Signal ◽

Diphenyltetrazolium Bromide ◽

Marine Compound

In this study, we aimed to illustrate the potential bio-effects of 3-bromo-4,5-dihydroxybenzaldehyde (3-BDB) on the antioxidant/cytoprotective enzyme heme oxygenase-1 (HO-1) in keratinocytes. The antioxidant effects of 3-BDB were examined via reverse transcription PCR, Western blotting, HO-1 activity assay, and immunocytochemistry. Chromatin immunoprecipitation analysis was performed to test for nuclear factor erythroid 2-related factor 2 (Nrf2) binding to the antioxidant response element of the HO-1 promoter. Furthermore, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that the cytoprotective effects of 3-BDB were mediated by the activation of extracellular signal-regulated kinase (ERK) and protein kinase B (PKB, Akt) signaling. Moreover, 3-BDB induced the phosphorylation of ERK and Akt, while inhibitors of ERK and Akt abrogated the 3-BDB-enhanced levels of HO-1 and Nrf2. Finally, 3-BDB protected cells from H2O2- and UVB-induced oxidative damage. This 3-BDB-mediated cytoprotection was suppressed by inhibitors of HO-1, ERK, and Akt. The present results indicate that 3-BDB activated Nrf2 signaling cascades in keratinocytes, which was mediated by ERK and Akt, upregulated HO-1, and induced cytoprotective effects against oxidative stress.

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