Molecular Mapping of Hydrogen Sulfide Targets in Normal Human Keratinocytes
Abstract
:1. Introduction
2. Results
2.1. NaHS Induces a Rapid and Transient Increase in H2S in Human Keratinocyte Cultures
2.2. NaHS Impairs the Growth of Human Keratinocyte Progenitors and Stem Cells
2.3. NaHS Stimulates the Synthesis of Superoxide Dismutase 2 (SOD2) by Cultured Human Keratinocytes
2.4. NaHS Exerts Specific Effects on the RNA Profile of Cultured Human Keratinocytes
2.5. NaHS Modifies the Secretary Profile of Cultured Human Keratinocytes
2.6. Distinct Molecular Pathways Mediate the Stimulating Effects of NaHS on the Synthesis of IL-1β and IL-18 by Cultured Human Keratinocytes
3. Discussion
4. Materials and Methods
4.1. Ethical Statement
4.2. Generation of Cultured Human Epidermal Sheets
4.3. NaHS
4.4. Measurements of H2S in Culture Supernatants
4.5. Fluorescence-Based Detection of Intracellular H2S
4.6. Clonogenic Assay
4.7. RNA–seq and Proteomics Analyses
4.8. Enzyme-Linked Immunosorbent Assay (ELISA)
4.9. Western Blot Analysis
4.10. Statistical Analysis
4.11. Data Availability
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
NAHS | Sodium hydrosulfide |
H2S | Hydrogen sulfide |
IL | Interleukin |
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GO Term | Adjusted p-Value |
---|---|
response to reactive oxygen species | 5.79 × 10−19 |
extrinsic apoptotic signaling pathway | 9.14 × 10−18 |
activation of cysteine-type endopeptidase activity involved in apoptotic process | 3.08 × 10−17 |
cellular response to reactive oxygen species | 6.10 × 10−17 |
glutathione metabolic process | 7.22 × 10−16 |
negative regulation of extrinsic apoptotic signaling pathway | 2.05 × 10−15 |
cellular response to mechanical stimulus | 1.00 × 10−14 |
I-kappaB kinase/NF-kappaB signaling | 2.73 × 10−13 |
interleukin-1-mediated signaling pathway | 2.73 × 10−13 |
toll-like receptor signaling pathway | 3.46 × 10−13 |
Gene Symbol | Gene Name |
---|---|
Up-regulated genes | |
CASP1 | Caspase-1 |
CAST | Calpastatin |
CUL3 | Cullin 3 |
CXCL2 | C-X-C motif chemokine ligand 2 |
GDF15 | Growth differentiation factor 15 |
NQO1 | NAD(P)H quinone dehydrogenase 1 |
SLC3A2 | Solute carrier family 3 member 2 |
UGT1A6 | UDP glucuronosyltransferase family 1 member A6 |
Down-regulated genes | |
CAMKK2 | Calcium/calmodulin dependent protein kinase kinase 2 |
HOMER3 | Homer scaffold protein 3 |
PTK2 | Protein tyrosine kinase 2 |
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Gross-Amat, O.; Guillen, M.; Gimeno, J.-P.; Salzet, M.; Lebonvallet, N.; Misery, L.; Auxenfans, C.; Nataf, S. Molecular Mapping of Hydrogen Sulfide Targets in Normal Human Keratinocytes. Int. J. Mol. Sci. 2020, 21, 4648. https://doi.org/10.3390/ijms21134648
Gross-Amat O, Guillen M, Gimeno J-P, Salzet M, Lebonvallet N, Misery L, Auxenfans C, Nataf S. Molecular Mapping of Hydrogen Sulfide Targets in Normal Human Keratinocytes. International Journal of Molecular Sciences. 2020; 21(13):4648. https://doi.org/10.3390/ijms21134648
Chicago/Turabian StyleGross-Amat, Olivia, Marine Guillen, Jean-Pascal Gimeno, Michel Salzet, Nicolas Lebonvallet, Laurent Misery, Céline Auxenfans, and Serge Nataf. 2020. "Molecular Mapping of Hydrogen Sulfide Targets in Normal Human Keratinocytes" International Journal of Molecular Sciences 21, no. 13: 4648. https://doi.org/10.3390/ijms21134648