MSF Enhances Human Antimicrobial Peptide β-Defensin (HBD2 and HBD3) Expression and Attenuates Inflammation via the NF-κB and p38 Signaling Pathways
Abstract
:1. Introduction
2. Results
2.1. MSF Significantly Enhanced the Expression of Human β-Defensin-2 and β-Defensin-3
2.2. Effect of MSF on the PI3K, NF-κB, and MAPKs Signaling Pathways in HaCaT Cells
2.3. MSF Promoted the Expression of HBD2 and HBD3 through p38 Pathway
2.4. NF-κB Involved in the Induction of HBD3 and HBD2 Expression
2.5. MSF Inhibited S. aureus-Induced Inflammation in THP1 Cells
2.6. TLR2 Was Involved in Regulating Human β-Defensin and Inflammation of MSF
2.7. MSF Ameliorated S. Aureus-Induced Skin Inflammation in Mice
3. Discussion
4. Materials and Methods
4.1. Preparation of MSF
4.2. Cell Culture and Stimulation
4.3. S. aureus Preparation
4.4. Cell Viability
4.5. qRT-PCR Analysis
4.6. Western Blot Analysis
4.7. Animal Experiments
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
Abbreviations
AKT | Protein kinase B |
AMPs | Antimicrobial peptides |
CCARM | Culture Collection of Antimicrobial Resistant Microbes |
DMEM | Dulbecco’s Modified Eagle’s Medium |
DMSO | Dimethyl sulfoxide |
ERK1/2 | Extracellular signal-regulated kinases 1 and 2 |
FBS | Fetal bovine serum |
FGE | F. glaberrima extracts |
hBD1 | Human β-defensins 1 |
hBD2 | Human β-defensins 2 |
hBD3 | Human β-defensins 3 |
HNP | Human neutrophil peptide |
IL-1β | Interleukin 1 beta |
IL-6 | Interleukin 6 |
IL-8 | Interleukin 8 |
IκB | Inhibitory kappa B alpha |
JNK | c-Jun N-terminal kinase |
MAPK | Mitogen-activated protein kinase |
MSF | Miracle synergy material made using F. glaberrima |
NF-κB | Nuclear Factor-kappa B |
PBS | Phosphate-buffered saline |
PI3K | Phosphoinositide 3-kinase |
TLR2 | Toll-like receptor 2 |
TNF-α | Tumor necrosis factor alpha |
TSB | Tryptic soy broth |
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Primers | Forward | Reverse |
---|---|---|
hBD1 | GTCGCCATGAGAACTTCCTACC | CATTGCCCTCCACTGCTGAC |
hBD2 | CCTGTTACCTGCCTTAAGAGTG | GAATCCGCATCAGCCACAG |
hBD3 | CTTCTGTTTGCTTTGCTCTTCC | CACTTGCCGATCTGTTCCTC |
hTNF-α | TGAGCACTGAAAGCATGATCC | ATCACTCCAAAGTGCAGCAG |
hIL-1β | TCTTTGAAGAAGAGCCCGTCCTC | GGATCCACACTCTCCAGCTGCA |
hIL-6 | CCTGAACCTTCCAAAGATGGC | CACCAGGCAAGTCTCCTCATT |
hIL-8 | TCTGTGTGAAGGTGCAGTTTT | GGGGTGGAAAGGTTTGGAGTA |
hTLR2 | TGTCTTGTGACCGCAATGGT | GTTGGACAGGTCAAGGCTTT |
hTLR4 | CCCTGAGGCATTTAGGCAGCTA | AGGTAGAGAGGTGGCTTAGGCT |
hGAPDH | GTGAAGGTCGGAGTCAACG | TGAGGTCAATGAAGGGGTC |
Antibody | Source | Category No. |
---|---|---|
p-PI3K | Cell Signaling Technology | 4257S |
PI3K | Cell Signaling Technology | 4228S |
p-AKT | Cell Signaling Technology | 9271S |
AKT | Cell Signaling Technology | 9272S |
p-NF-κB | Cell Signaling Technology | 3033S |
NF-κB | Santa Cruz Biotechnology | sc-8008 |
p-p38 | Cell Signaling Technology | 9211S |
p38 | Santa Cruz Biotechnology | sc-535 |
p-ERK1/2 | Cell Signaling Technology | 9101S |
ERK1/2 | Cell Signaling Technology | 9102S |
p-JNK | Santa Cruz Biotechnology | sc-6254 |
JNK | Santa Cruz Biotechnology | sc-7345 |
HBD2 | Abcam | ab9871 |
HBD3 | Abcam | ab172703 |
GAPDH | Cell Signaling Technology | 14C10 |
Left Ear | Right Ear | |||
---|---|---|---|---|
Compound | Pathogen | Compound | Pathogen | |
Group 1 | None | None | PBS | None |
Group 2 | None | None | None | S. aureus |
Group 3 | K8 lysate | None | PBS | None |
Group 4 | K8 lysate | S. aureus | PBS | S. aureus |
Group 5 | MSF | None | PBS | None |
Group 6 | MSF | S. aureus | PBS | S. aureus |
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Nguyen, A.-T.; Kim, M.; Kim, Y.-E.; Kim, H.; Lee, S.; Lee, Y.; Kim, K.-Y. MSF Enhances Human Antimicrobial Peptide β-Defensin (HBD2 and HBD3) Expression and Attenuates Inflammation via the NF-κB and p38 Signaling Pathways. Molecules 2023, 28, 2744. https://doi.org/10.3390/molecules28062744
Nguyen A-T, Kim M, Kim Y-E, Kim H, Lee S, Lee Y, Kim K-Y. MSF Enhances Human Antimicrobial Peptide β-Defensin (HBD2 and HBD3) Expression and Attenuates Inflammation via the NF-κB and p38 Signaling Pathways. Molecules. 2023; 28(6):2744. https://doi.org/10.3390/molecules28062744
Chicago/Turabian StyleNguyen, Anh-Thu, Minho Kim, Ye-Eun Kim, Hangeun Kim, Sanghyun Lee, Yunji Lee, and Ki-Young Kim. 2023. "MSF Enhances Human Antimicrobial Peptide β-Defensin (HBD2 and HBD3) Expression and Attenuates Inflammation via the NF-κB and p38 Signaling Pathways" Molecules 28, no. 6: 2744. https://doi.org/10.3390/molecules28062744