Dataset Information

CBG and EAA for melanocytes protection

ABSTRACT: Aims: This study aims to assess the effect of simultaneous use of cannabigerol (CBG) and 3-O-ethyl ascorbic acid (EAA) on changes in the skin melanocyte proteome induced by UVA radiation, with particular emphasis on modifications in the structure of metabolically important proteins. Results: Proteomic analysis allowed to identify 1248 proteins with statistically significantly changed expression following melanocytes irradiation and/or incubation with CBG/EAA. The top 25 proteins with most strongly modified expression included mainly proteins involved in cells protection/antioxidant response, as well as pro-inflammatory and proapoptotic signalization. Moreover, in melanocytes irradiated with UVA, the levels of lipid peroxidation product, 4-hydroxynonenal (4-HNE) and its protein adducts increase, as well as significant changes in the profile of proteins modified by 4-HNE are observed. CBG and EAA, especially when used together, largely reverse this effect. Innovation: This study for the first time shows the combined effect of CBG and EAA on the proteome of melanocytes after their exposure to UVA radiation, that applies to both changes in protein expression and intracellular signaling based on protein modification by lipid peroxidation products. Conclusion: It can be summarized that CBG and EAA can provide melanocytes with the most effective protection against oxidative stress, and perhaps even protect the skin structure against carcinogenesis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Melanocyte, Cell Culture

SUBMITTER: Agnieszka Gęgotek

LAB HEAD: Agnieszka Gęgotek

PROVIDER: PXD074276 | Pride | 2026-06-15

REPOSITORIES: Pride

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Publications

Proteomic Analysis of the Metabolic Response of UVA-Exposed Melanocytes Following Co-Treatment with Cannabigerol and 3-<i>O</i>-Ethylascorbic Acid.

Mucha Magda M Ryšavá Alena A Jarocka-Karpowicz Iwona I Maruška Audrius A Skrzydlewska Elżbieta E Gęgotek Agnieszka A

Cells 20260523 11

The aim of this study was to analyze the effect of concomitant use of cannabigerol (CBG) and 3-<i>O</i>-ethylascorbic acid (EAA) on changes in the proteome of UVA-irradiated skin melanocytes, with particular emphasis on adduct formation between lipid peroxidation products and metabolically important proteins. Proteomic analysis allowed the identification of 1248 proteins with statistically significantly changed expression following melanocytes irradiation and/or incubation with CBG/EAA. The top ...[more]

PMID: 42274558

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Project description:Lipid extracts, specifically from marine microalgae Nannochloropsis oceanica (N.o.) and freshwater microalgae Chlorococcum amblystomatis (C.a.) in this research proposal, offer a great potential in the development of new pharmacotherapeutic applications against skin disease, accompanied by oxidative stress and associated chronic inflammation, and even skin cancer, due to their potential antioxidant, anti-inflammatory and anti-cancer properties reported in the literature. Therefore, lipid extracts obtained from two species of microalgae of different origins, characterized by a high content of bioactive compounds of a lipophilic nature, may also have cytoprotective effects through the modification of antioxidant and anti-inflammatory proteins, and ultimately the associated alterations in the intracellular molecular signaling via the changes in both associated protein expression, structure, and functionality. Thus, a proteomic research, using treatment with lipid extracts obtained from N.o. and C.a., has been planned to perform for analyzing the changes in the whole proteomic profile of human skin fibroblasts – in which its proteome is known to be dramatically disturbed by UVA radiation and plays a critical role in the intracellular signal transduction regarding important biological processes, such as the redox-dependent inflammatory response – under oxidative stress condition resulted by UVA irradiation. In this research, the proteins involved in the regulation of redox balance and inflammatory response will be particularly analyzed to understand the molecular mechanism behind the cytoprotective action of these microalgae lipid extracts. Moreover, a pull-down immunoprecipitation assay-combined proteomic analysis showing the human caspase-1 protein interactome area (associated with NLRP1, NLRP3, NLRC4, and AIM2 complexes) will be performed to understand the effect of these lipid extracts on the changes in the interaction between the intracellular redox signaling and inflammasome activities. The obtained proteomic data will indicate the regenerative/protective effect of lipid extracts on the protein homeostasis of fibroblasts exposed to UVA radiation, analyzed by assessing the molecular activity underlying the potential antioxidant and/or anti-inflammatory effects of these extracts. Also, analysis of the interaction area of inflammasome complexes may demonstrate the influence of these lipid extracts on the innate immune response of skin fibroblasts, which closely interacts with redox metabolism. This is another important problem in the development of effective antioxidant therapies, the limitations and failures of which are highlighted in clinical trials. Furthermore, the proteomic approach is a privileged analytical tool enabling knowledge and understanding of the communication between intracellular metabolic changes resulting from the interaction of oxidative stress and inflammatory protein networks in response to physicochemical extracellular factors.

Project description:Ultraviolet (UV) wavebands in sunlight are immunomodulatory. About half the amount of UVA within a minimum erythemal dose of sunlight is systemically immunosuppressive, while higher doses protect from UVB immunosuppression in mice. We have previously shown that these responses to UVA are genetically restricted as they occur in C57BL/6 but not Balb/c mice. We used gene set enrichment analysis of microarray data and real-time RT-PCR confirmation to determine the molecular mechanisms associated with UVA immunomodulation. We found up-regulation of mRNA for the alternative complement pathway. The core-enriched genes complement component 3, properdin and complement factor B were all activated by the immunosuppressive dose of UVA only in UVA-responsive C57BL/6 but not unresponsive BALB/c mice. This therefore matched the genetic restriction and dose responsiveness of UVA immunosuppression. The immune-protective higher UVA dose prevented UVB from down regulating chemokine receptor 7 and IL-12B, and decreased IL-10, supporting previous identification of IL-12 and IL-10 in high dose UVA protection from UVB immunosuppression. Our study has identified activation of the alternative complement pathway as a trigger of UVA-induced systemic immunosuppression and suggests that this pathway is likely to be an important sensor of UVA-induced damage to the skin. 24 hours after UVA, UVB and ssUV irradiation, a 1 cm2 uniform section of skin was excised from the dorsal surface of irradiated and control mice. Total RNA was then extracted from the whole skin using TRIzol reagent (Gibco Invitrogen Life Technologies, Carlsbad, CA, USA) according to the manufacturerâ??s instructions, purified, DNase treated and reverse transcribed into cDNA. For the microarray study a direct incorporation of Cyanine 3-dCTP and Cyanine 5-dCTP fluorescent dyes (Perkin Elmer Life Sciences, Inc. Boston, MA, USA) was used for cDNA synthesis. For each UV dose, a reference design was used to compare an unirradiated control against an irradiated sample. Microarray experiments used compugen 22k mouse oligonucleotide microarray slides (The Clive and Vera Ramaciotti Centre for Gene Function Analysis, Sydney Australia (http://www.ramaciotti.unsw.edu.au). Lower and higher UVA doses were used. C57BL/6 mice were irradiated with lower UVA, higher UVA, UVB, or ssUV; Balb/C mice were irradiated with lower or higher UVA. Experiments were replicated 6 times for each UV dose. A fluorescent dye swap was done for each alternate hybridisation to reduce systematic dye bias of incorporated fluorescent dyes.

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Dataset Information

CBG and EAA for melanocytes protection

Dataset's files

Publications

Proteomic Analysis of the Metabolic Response of UVA-Exposed Melanocytes Following Co-Treatment with Cannabigerol and 3-<i>O</i>-Ethylascorbic Acid.

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Dataset Information

CBG and EAA for melanocytes protection

Dataset's files

Publications

Proteomic Analysis of the Metabolic Response of UVA-Exposed Melanocytes Following Co-Treatment with Cannabigerol and 3-&lt;i&gt;O&lt;/i&gt;-Ethylascorbic Acid.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Proteomic Analysis of the Metabolic Response of UVA-Exposed Melanocytes Following Co-Treatment with Cannabigerol and 3-<i>O</i>-Ethylascorbic Acid.