The effects of UVB and the organic osmolyte betaine on rat epidermal keratinocytes in an organotypic culture system
Ontology highlight
ABSTRACT:
ORGANISM(S): Rattus norvegicus
PROVIDER: GSE63004 | GEO | 2015/10/26
SECONDARY ACCESSION(S): PRJNA266534
REPOSITORIES: GEO
Similar Datasets
Project description:Betaine (trimethylglycine) is a non-toxic, highly water-soluble organic osmolyte widely used in skin care due to its assumed moisturizing and protective properties, but only few studies have addressed its specific effects in skin. Here, the cellular and molecular targets of betaine were analyzed by genome-wide expression analysis in organotypic cultures of rat epidermal keratinocytes (REK). In this model, we also examined whether betaine modifies the impacts of acute UVB exposure. The expression of several genes relevant to epidermal biology, proliferation/differentiation or malignancy as well as solute transport were verified by independent methods (qRT-PCR, western blotting). The data concerning changes in calcium metabolism after UVB exposure has been published separately (Bart et al., Br. J. Dermatol. 171:376-387, 2014). Organotypic cultures of rat epidermal keratinocytes (REK) were prepared by plating cells on a collagen-coated insert (submerged in medium), lifting the confluent cell layer to the air-liquid interface 3 days after plating. These 3D cultures were divided in four treatment groups with three replicates in each: control, betaine (10 mM), UVB (30 mJ/cm2) and betaine + UVB. Betaine was added to the cultures for 11 days, starting from day 4 until sample collection. UVB exposure was performed 24 h prior to sample collection for 2-week-old 3D cultures with a well-formed epidermal layer. For further details on the experimental set-up and characteristics of the UV source, please refer to Bart et al. (Br. J. Dermatol. 171:376-387, 2014) and Rauhala et al. (J. Biol. Chem. 288:17999-18012, 2013).
2015-10-26 | E-GEOD-63004 | biostudies-arrayexpress
Project description:The effects of UVB and the organic osmolyte betaine on rat epidermal keratinocytes in an organotypic culture system
| PRJNA266534 | ENA
Project description:Betaine critically contributes to the control of hepatocellular hydration and provides protection of the liver from different kinds of stress. This study investigates to what extent hepatocellular hydration changes affect the expression levels of enzymes involved in the metabolism of betaine and related organic osmolytes by using qRT-PCR gene expression studies in rat hepatoma cells as well as metabolic and gene expression profiling in 5,10 - methylene tetrahydrofolate reductase (MTHFR) deficient primary hepatocytes. The results demonstrate a coordinated regulation of betaine degradation and synthesis under anisoosmotic conditions. Expression of betaine degrading enzymes is downregulated by hyperosmolarity and strongly induced by hypoosmolarity. In contrast, synthesis of glycerophosphocholine from phosphoethanolamine and conversion of choline to betaine are both induced by hyperosmolarity but decreased under hypoosmotic conditions. In addition we evaluated the flux of choline and its derivates in liver and plasma of methylene tetrahydrofolate reductase knockout (Mthfr-/-) mice by tandem mass spectrometry. Analyses of system-wide alterations of osmolyte metabolism with microarray studies revealed expression changes similar to those after hypoosmotic exposure in this betaine depletion model. In conclusion, regulation of betaine synthesis and degradation and concomitant changes in intracellular osmolyte concentrations contribute to long-term adaptation to anisoosmotic exposure of the liver. Expression of 280 genes were analyzed in wild type and mthr-/- mice (n=7) with spotted oligonucleotides.
2013-04-01 | E-GEOD-42275 | biostudies-arrayexpress
Project description:Background: Atopic dermatitis (AD) is a common inflammatory skin disease exhibiting a predominantly Th2/"T22" immune activation and a defective epidermal barrier. Narrow-band UVB (NB-UVB) is considered an efficient treatment for moderate to severe AD. In psoriasis, NB-UVB has been found to suppress the Th1/Th17 immune polarization with subsequent reversal of epidermal hyperplasia. The immunomodulatory effects of this treatment are largely unknown in AD. Our study evaluates the effects of NB-UVB on immune and barrier abnormalities in AD, aiming to establish reversibility of disease and biomarkers of therapeutic response. Methods: 12 moderate-to-severe chronic AD patients received NB-UVB phototherapy 3 times weekly for up to 12 weeks. Lesional and non-lesional skin biopsies were obtained before and after treatment and evaluated by gene-expression and immunohistochemistry studies. Results: All patients had at least a 50% reduction in SCORing of AD (SCORAD) index with NB-UVB phototherapy. The Th2, T22, and Th1 immune pathways were suppressed and measures of epidermal hyperplasia and differentiation also normalized after phototherapy. The reversal of disease activity was associated with elimination of inflammatory leukocytes, Th2/"T22"-associated cytokines and chemokines, and normalized expression of barrier proteins. Conclusions: Our study shows reversal of both the epidermal defects and underlying immune activation in AD. By determining the correlation of variables with therapeutic response, we have defined a set of biomarkers of disease response that associate resolved Th2 and T22 inflammation with reversal of barrier pathology. This data supports the "inside-out" hypothesis of AD, suggesting that it is a disease primarily driven by an immune stimulus. genomic profiling of treatment effect of NB-UVB in AD in both lesional and non-lesional AD skin from 10 patients. Treatment effect, disease state analysis
2011-07-01 | E-GEOD-27887 | biostudies-arrayexpress
Project description:Betaine critically contributes to the control of hepatocellular hydration and provides protection of the liver from different kinds of stress. This study investigates to what extent hepatocellular hydration changes affect the expression levels of enzymes involved in the metabolism of betaine and related organic osmolytes by using qRT-PCR gene expression studies in rat hepatoma cells as well as metabolic and gene expression profiling in 5,10 - methylene tetrahydrofolate reductase (MTHFR) deficient primary hepatocytes. The results demonstrate a coordinated regulation of betaine degradation and synthesis under anisoosmotic conditions. Expression of betaine degrading enzymes is downregulated by hyperosmolarity and strongly induced by hypoosmolarity. In contrast, synthesis of glycerophosphocholine from phosphoethanolamine and conversion of choline to betaine are both induced by hyperosmolarity but decreased under hypoosmotic conditions. In addition we evaluated the flux of choline and its derivates in liver and plasma of methylene tetrahydrofolate reductase knockout (Mthfr-/-) mice by tandem mass spectrometry. Analyses of system-wide alterations of osmolyte metabolism with microarray studies revealed expression changes similar to those after hypoosmotic exposure in this betaine depletion model. In conclusion, regulation of betaine synthesis and degradation and concomitant changes in intracellular osmolyte concentrations contribute to long-term adaptation to anisoosmotic exposure of the liver.
2013-04-01 | GSE42275 | GEO
Project description:Proline betaine, and to a much lesser extent, N-methyl L-proline, are found in citrus fruits and often present in the human metabolome. However, most of the N-methyl L-proline found in an animal model metabolome is produced by the microbiota, yet organisms and enzymes capable of producing N-methyl proline in the anoxic intestine have been unknown. The anaerobe Eubacterium limosum ATCC 8486 makes acetate and butyrate from various substrates and is found in the human intestine. We found that this organism demethylates proline betaine and excretes N-methyl proline during growth. The proteome of proline betaine-grown Eubacterium limosum was obtained in order to identify enzymes required for growth on proline, in particular to identify components that are unique to growth on proline in comparison to other substrates for acetogenesis, such as lactic acid. Comparison of the proteomes of the bacteria on proline betaine and lactic acid led to identification of the proteins involved in a proline betaine:tetrahydrofolate methyltransferase system which was biochemically verified. The key proline betaine demethylating enzyme is a member of the widespread TMA methyltransferase protein superfamily.
2019-07-25 | PXD013962 | Pride
Project description:Phototherapy is an effective therapy and may induce remission of psoriasis. Previous studies have established the action spectrum of clearance and that apoptosis is differentially induced in psoriasis plaques by clinically effective wavelengths of ultraviolet B (UVB). The aim of this study was to investigate the molecular mechanisms regulating psoriasis plaque resolution by studying the transcriptomic response to clinically effective (311nm, narrow band) UVB compared to a clinically ineffective (290nm) wavelength. We irradiated lesional psoriatic skin in vivo with a single 3 MED (minimal erythemal dose) of 311nm or 290nm wavelength of UVB and performed skin biopsies at 4h or 18h post irradiation and from un-irradiated lesional skin. Forty-eight micro-dissected epidermal samples were analysed using the Illumina DASL array platform from 20 psoriatic patients. Bioinformatic analysis identified differentially expressed genes (DEGs) associated with 311nm but not 290nm irradiation; these DEGs were subject to Ingenuity pathway and upstream regulator analysis. The number of differentially regulated epidermal genes was greatest at 18h following UVB, after irradiation with clinically effective (311nm) UVB. The main pathways differentially affected by 311nm UVB only were apoptosis, necrosis, acute phase signalling, p53 signalling and chemotaxis. The greatest fold change observed was a 7.5 fold increase in expression of CDKN1A (WAF1/ p21), the p53 target gene, following irradiation with 311nm UVB but not 290nm (clinically ineffective UVB). Acute phase, LXR and PTEN signalling, dendritic cell maturation, granulocyte adhesion and atherosclerotic pathways were also differentially regulated by 311nm compared to 290nm UVB. This work provides insight into the molecular mechanisms regulating psoriatic remodelling in response to UV phototherapy, supports a key role for apoptosis and cell death in psoriasis plaque clearance, and identifies a number of novel therapeutic pathways. Further studies may lead to development of potential biomarkers to assess which patients are more likely to respond to UVB.
2021-04-21 | GSE162998 | GEO
Project description:Purpose: Understanding the mechanism of action of lauryl betaine against Cryptococcus neoformans and Malassezia restricta. Methods: The transcriptome profile of the lauryl betaine-treated cells compared to that of untreated cells were generated by RNA-seq Illumina Hiseq. Results: A number of genes were differentially expressed in the lauryl betaine-treated cells, which include genes involved in ergosterol biosynthetic pathway.
2019-08-22 | GSE124734 | GEO
Project description:Background: Atopic dermatitis (AD) is a common inflammatory skin disease exhibiting a predominantly Th2/“T22” immune activation and a defective epidermal barrier. Narrow-band UVB (NB-UVB) is considered an efficient treatment for moderate to severe AD. In psoriasis, NB-UVB has been found to suppress the Th1/Th17 immune polarization with subsequent reversal of epidermal hyperplasia. The immunomodulatory effects of this treatment are largely unknown in AD. Our study evaluates the effects of NB-UVB on immune and barrier abnormalities in AD, aiming to establish reversibility of disease and biomarkers of therapeutic response. Methods: 12 moderate-to-severe chronic AD patients received NB-UVB phototherapy 3 times weekly for up to 12 weeks. Lesional and non-lesional skin biopsies were obtained before and after treatment and evaluated by gene-expression and immunohistochemistry studies. Results: All patients had at least a 50% reduction in SCORing of AD (SCORAD) index with NB-UVB phototherapy. The Th2, T22, and Th1 immune pathways were suppressed and measures of epidermal hyperplasia and differentiation also normalized after phototherapy. The reversal of disease activity was associated with elimination of inflammatory leukocytes, Th2/“T22”-associated cytokines and chemokines, and normalized expression of barrier proteins. Conclusions: Our study shows reversal of both the epidermal defects and underlying immune activation in AD. By determining the correlation of variables with therapeutic response, we have defined a set of biomarkers of disease response that associate resolved Th2 and T22 inflammation with reversal of barrier pathology. This data supports the “inside-out” hypothesis of AD, suggesting that it is a disease primarily driven by an immune stimulus.
2011-07-01 | GSE27887 | GEO
Project description:In order to explore the effect of betaine on NAFLD and its mechanism, we divided mice into high-fat diet (HF) and high-fat diet with betaine (HB) groups, and used N6-methyladenosine sequence (m6A-seq), RNA-seq and mRNAs bound to ribosome-nascent chain complex sequencing (RNC-seq) to evaluate the gene expression levels in the liver of each group.
2021-05-20 | GSE174550 | GEO