A Small Molecule, 4-Phenylbutyric Acid, Suppresses HCV Replication via Epigenetically Induced Hepatic Hepcidin
Abstract
:1. Introduction
2. Results
2.1. 4-PBA Inhibited HCV RNA Replication
2.2. 4-PBA Induced the Expression of the Antimicrobial Peptide (AMP), Hepcidin
2.3. 4-PBA Enhanced the IFN-α Response
2.4. Hepcidin Mediated 4-PBA-Induced IFN-α Signaling
2.5. 4-PBA Disrupted MW Formation
2.6. 4-PBA Inhibited the Formation of Double Membranous Vesicles (DMVs)
2.7. 4-PBA Interfered with Molecular Interactions in the HCV RNA Replication Complex
2.8. The Anti-HCV Effects of 4-PBA in a Mouse Model
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Reagents
4.3. Quantitative Real-Time Reverse Transcription Polymerase Chain Reaction (qRT-PCR)
4.4. Determination of Cell Viability
4.5. Chromatin Immunoprecipitation Assay
4.6. Membrane Flotation Assay
4.7. Luciferase Assay
4.8. Immunoblotting
4.9. Immunofluorescence Analysis
4.10. Immunoprecipitation
4.11. Immunohistochemistry
4.12. DNAs and siRNAs Transfection
4.13. Electron Microscopy
4.14. Animal Procedures
4.15. Data and Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
HCV | Hepatitis C Virus |
4-PBA | 4-Phenyl butyrate |
HDACi | Histone Deacetylase Inhibitor |
HCC | Hepatocellcular Carcinoma |
DAAs | Direct Acting antiviral reagents |
FDA | Food and Drug Administration |
ER | Endoplasmic Reticulum |
AMPs | Antimicrobial Peptides |
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Gene | Direction | Sequence |
---|---|---|
HCV | forward | CGGGAGAGCCATAGTGGTCTG CG |
reverse | CTCGCAAGCACCCTATCA GGCAGTA | |
b-actin | forward | AGGCTGTGCTGTCCCTCT |
reverse | TCCGGTGAGGAGGATGCG | |
hepcidin | forward | ACCAGAGCAAGCTCAAGACC |
reverse | CAGGGCAGGTAGGTTCTACG | |
IFN-a | forward | TTTCTCCTGCCTGAAGGACAG |
reverse | GCTCATGATTTCTGCTCTGACA | |
OAS1 | forward | AGGTGGTAAAGGGTGGCTCC |
reverse | ACAACCAGGTCAGCGTCAGAT | |
IFIT1 | forward | TGGCTAAGCAAAACCCTGCA |
reverse | TCTGGCCTTTCAGGTGTTTCAC | |
IRF1 | forward | CTCACCAGGAACCAGAGGAA |
reverse | TGAGTGGTGTAACTGCTGTGG | |
IRF3 | forward | ACCAGCCGTGGACCAAGAG |
reverse | TACCAAGGCCCTGAGGCAC | |
IRF7 | forward | TGGTCCTGGTGAAGCTGGAA |
reverse | GATGTCGTCATAGAGGCTGTTGG | |
PKR | forward | TCTCTGGCGGTCTTCAGAAT |
reverse | ACTCCCTGCTTCTGACGGTA | |
hVAP-33 | forward | GAAGACTACAGCACCTCGCC |
reverse | GCCTCTTTCCACACAGCTTC | |
Hepcidin_locus | forward | CTGTTTTCCCACAACAGGTG |
reverse | CTCAGTGCTCGGGTGTCTC |
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Kim, K.; Lee, Y.-s.; Jeong, S.; Kim, D.; Chon, S.; Pak, Y.K.; Kim, S.; Ha, J.; Kang, I.; Choe, W. A Small Molecule, 4-Phenylbutyric Acid, Suppresses HCV Replication via Epigenetically Induced Hepatic Hepcidin. Int. J. Mol. Sci. 2020, 21, 5516. https://doi.org/10.3390/ijms21155516
Kim K, Lee Y-s, Jeong S, Kim D, Chon S, Pak YK, Kim S, Ha J, Kang I, Choe W. A Small Molecule, 4-Phenylbutyric Acid, Suppresses HCV Replication via Epigenetically Induced Hepatic Hepcidin. International Journal of Molecular Sciences. 2020; 21(15):5516. https://doi.org/10.3390/ijms21155516
Chicago/Turabian StyleKim, Kiyoon, Young-seok Lee, Suyun Jeong, Daehong Kim, Suk Chon, Youngmi Kim Pak, Sungsoo Kim, Joohun Ha, Insug Kang, and Wonchae Choe. 2020. "A Small Molecule, 4-Phenylbutyric Acid, Suppresses HCV Replication via Epigenetically Induced Hepatic Hepcidin" International Journal of Molecular Sciences 21, no. 15: 5516. https://doi.org/10.3390/ijms21155516