Inhibition of HDAC Enzymes Contributes to Differential Expression of Pro-Inflammatory Proteins in the TLR-4 Signaling Cascade
Abstract
:1. Introduction
2. Results
2.1. Effects of HDAC Inhibitors on Cell Proliferation and Apoptosis Induction
2.2. Impact of TSA and SAHA on the Expression of Pro-Inflammatory Genes
2.3. TSA and SAHA Enhance LPS-Induced NF-κB Activation
2.4. Effects of TSA and SAHA Are Only Partially Mediated by Histone Modulations
3. Discussion
4. Materials and Methods
4.1. Drugs
4.2. Cells
4.3. siRNA
4.4. Sulforhodamine B Cytotoxicity Assay
4.5. TUNEL Staining
4.6. Polymerase Chain Reactions (PCR)
COX-2 | FW 5’-AGACACTCAGGTAGACATGATCTACCCT-3′ |
RV 5′-GGCACCAGACCAAAGACTTCC-3′ | |
IKKε | FW 5′-GTACAAGGCCCGAAACAAGA-3′ |
RV 5′-TCCTCCACTGCGAATAGCTT-3′ | |
IL-1β | FW 5′-CTGGTGTGTGACGTTCCCATTA-3′ |
RV 5′-CCGACAGCACGAGGCTTT-3′ | |
IL-10 | FW 5′-GCTCTTACTGACTGGCATGAG-3′ |
RV 5′-CGCAGCTCTAGGAGCATGTG-3′ | |
iNOS | FW 5′-ACACAGCGCTACAACATCCT-3′ |
RV 5′-TGGAGCACAGCCACATTGAT-3′ | |
TNFα | FW 5′-GCTGAGCTCAAACCCTGGTA-3′ |
RV 5′-CGGACTCCGCAAAGTCTAAG-3′ | |
GAPDH | FW 5′-CAATGTGTCCGTCGTGGATCT-3′ |
RV 5′-GTCCTCAGTGTAGCCCAAGATG-3′ |
4.7. ChIP Assay
- FW
- 5′-GATCGACAAGTGCAATCCCC-3′
- RV
- 5′-AGCCACTCAAATGCACTGTG-3′
4.8. Western Blot Analysis
4.9. p65 TransAM ELISA
4.10. Flow Cytometry
4.11. Data Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
AcH3 | Acetyl histone H3 |
AmD | Actinomycin D |
BMM | Bone marrow macrophages |
ChIP | Chromatin immunoprecipitation |
COX-2 | Cyclooxygenase 2 |
DMSO | Dimethylsulfoxide |
ELISA | Enzyme linked immunosorbent assay |
FACS | Fluorescence activated cell sorting |
FCS | Fetal calf serum |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
HAT | Histone acetylase |
HDAC | Histone deacetylase |
HDACi | Histone deacetylase inhibitor |
IFN | Interferone |
IκBa | Inhibitor kappa B protein alpha |
IKKε | I-κB kinase epsilon |
IL | Interleukin |
LPS | lipopolysaccharide |
M-CSF | murine macrophage colony stimulating factor |
iNOS | Inducible nitric oxide synthase |
NFκB | Nuclear factor kappa B |
PCNA | Proliferating-Cell-Nuclear-Antigen |
RLP30 | Receptor-like protein 30 |
qRT-PCR | Quantitative reverse transcription polymerase chain reaction |
TCA | Trichloroacetic acid |
TLR | Toll-like receptor |
TNF | Tumor necrosis factor |
TSA | Trichostatin A |
TUNEL | TdT-mediated dUTP nick end labeling |
SAHA | Suberoylanilide hydroxamic acid |
SRB | Sulforhodamine B |
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Weiss, U.; Möller, M.; Husseini, S.A.; Manderscheid, C.; Häusler, J.; Geisslinger, G.; Niederberger, E. Inhibition of HDAC Enzymes Contributes to Differential Expression of Pro-Inflammatory Proteins in the TLR-4 Signaling Cascade. Int. J. Mol. Sci. 2020, 21, 8943. https://doi.org/10.3390/ijms21238943
Weiss U, Möller M, Husseini SA, Manderscheid C, Häusler J, Geisslinger G, Niederberger E. Inhibition of HDAC Enzymes Contributes to Differential Expression of Pro-Inflammatory Proteins in the TLR-4 Signaling Cascade. International Journal of Molecular Sciences. 2020; 21(23):8943. https://doi.org/10.3390/ijms21238943
Chicago/Turabian StyleWeiss, Ulrike, Moritz Möller, Sayed Adham Husseini, Christine Manderscheid, Julia Häusler, Gerd Geisslinger, and Ellen Niederberger. 2020. "Inhibition of HDAC Enzymes Contributes to Differential Expression of Pro-Inflammatory Proteins in the TLR-4 Signaling Cascade" International Journal of Molecular Sciences 21, no. 23: 8943. https://doi.org/10.3390/ijms21238943