MicroRNA-27a-5p Downregulates Expression of Proinflammatory Cytokines in Lipopolysaccharide-Stimulated Human Dental Pulp Cells via the NF-κB Signaling Pathway
Abstract
:1. Introduction
2. Results
2.1. LPS/NF-κB Signaling Promotes miR-27a-5p Expression and Proinflammatory Mediator Levels in LPS-Stimulated hDPCs
2.2. MiR-27a-5p Downregulates Synthesis of Proinflammatory Mediators via the NF-κB Pathway in LPS-Stimulated hDPCs
2.3. MiR-27a-5p Targets TAB1, IRAK4, and RELA in NF-κB Signaling
2.4. MiR-27a-5p Exerts a Downregulatory Effect on NF-κB Activity and Proinflammatory Cytokine Expression in LPS-Stimulated Rat Pulp Tissue Ex Vivo
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatments
4.2. Establishment of the Pulpitis Model in Rats
4.3. Ex Vivo miR-27a Mimic Transfection
4.4. Reverse Transcription-Quantitative Polymerase Chain Reaction
4.5. Western Blotting
4.6. Cytometric Bead Array
4.7. Luciferase Assays Using NF-κB and TAB1 3′-UTR Reporter Vectors
4.8. Immunofluorescence
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Forward | Reverse | Accession No. | Size, bp |
---|---|---|---|---|
<human> | ||||
ACTB | 5′-GTAGCACAGCTTCTCCTTAATGTCA-3′ | 5′-CTGACTGACTACCTCATGAAGATCC-3′ | NM_001101.3 | 102 |
IL6 | 5′-TATACCTCAAACTCCAAAAGACCAG-3′ | 5′-ACAAGAGTAACATGTGTGAAAGCAG-3′ | NM_000600.4 | 157 |
IL8 | 5′-TCAGTGCATAAAGACATACTCCAAA-3′ | 5′-TCTTCCATCAGAAAGCTTTACAATAA-3′ | NM_000584.4 | 121 |
MCP1 | 5′-CACCTGCTGTTATAACTTCACCAAT-3′ | 5′-GTTGAAAGATGATAAGCCCACTCTA-3′ | NM_002982.4 | 130 |
TAB1 | 5′-ATCCCTCAGTGCCAACTAAACC-3′ | 5′-GAAGATCCCAGTGCACAAGTCA-3′ | NM_153497.3 | 137 |
IRAK4 | 5′-CGGAAATCTCTTTATCATCCGTGAG-3′ | 5′-GCACATATGTTGATGGTGTTATGGG-3′ | NM_001351341.2 | 126 |
RELA | 5′-TTCCAAGTTCCTATAGAAGAGCAGC-3′ | 5′-TCAAAGATGGGATGAGAAAGGACAG-3′ | NM_021975.4 | 134 |
FSTL1 | 5′-CCATGACCTGTGACGGAAAGAAT-3′ | 5′-TTAGATCTCTTTGGTGCTCACTCT-3′ | NM_007085.5 | 137 |
<rat> | ||||
Actb | 5′-GTAAAGACCTCTATGCCAACACAGT-3′ | 5′-GGAGCAATGATCTTGATCTTCATGG -3′ | NM_031144.3 | 127 |
Il6 | 5′-TAAGGACCAAGACCATCCAACTCAT-3′ | 5′-AGTGAGGAATGTCCACAAACTGATA-3′ | NM_012589.2 | 125 |
Mcp1 | 5′-CTAAGGACTTCAGCACCTTTGAATG-3′ | 5′-GTTCTCTGTCATACTGGTCACTTCT-3′ | NM_031530.1 | 120 |
Tab1 | 5′-TAGTGTCTGCTTCTGTTAGATCCTG-3′ | 5′-AATCAGCTTCCTCATCAGAGTGAAA-3′ | NM_001109976.2 | 134 |
Irak4 | 5′-CTGAACGTGCTTTGTCTTTAACAAG-3′ | 5′-GTTGAAATGAGCTCCGTACTAAGTG-3′ | NM_001106791.2 | 121 |
Rela | 5′-CTTTCTCAAGTGCCTTAATAGCAGG-3′ | 5′-TTCAGAGCTAGAAAGAGCAAGAGTC-3′ | NM_199267.2 | 121 |
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Wang, S.; Kawashima, N.; Han, P.; Sunada-Nara, K.; Yu, Z.; Tazawa, K.; Fujii, M.; Kieu, T.Q.; Okiji, T. MicroRNA-27a-5p Downregulates Expression of Proinflammatory Cytokines in Lipopolysaccharide-Stimulated Human Dental Pulp Cells via the NF-κB Signaling Pathway. Int. J. Mol. Sci. 2024, 25, 9694. https://doi.org/10.3390/ijms25179694
Wang S, Kawashima N, Han P, Sunada-Nara K, Yu Z, Tazawa K, Fujii M, Kieu TQ, Okiji T. MicroRNA-27a-5p Downregulates Expression of Proinflammatory Cytokines in Lipopolysaccharide-Stimulated Human Dental Pulp Cells via the NF-κB Signaling Pathway. International Journal of Molecular Sciences. 2024; 25(17):9694. https://doi.org/10.3390/ijms25179694
Chicago/Turabian StyleWang, Shihan, Nobuyuki Kawashima, Peifeng Han, Keisuke Sunada-Nara, Ziniu Yu, Kento Tazawa, Mayuko Fujii, Thoai Quoc Kieu, and Takashi Okiji. 2024. "MicroRNA-27a-5p Downregulates Expression of Proinflammatory Cytokines in Lipopolysaccharide-Stimulated Human Dental Pulp Cells via the NF-κB Signaling Pathway" International Journal of Molecular Sciences 25, no. 17: 9694. https://doi.org/10.3390/ijms25179694