Porphyromonas gingivalis Outer Membrane Vesicles Stimulate Gingival Epithelial Cells to Induce Pro-Inflammatory Cytokines via the MAPK and STING Pathways
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Cultures
2.2. OMV Isolation
2.3. Endotoxin Activity Assay
2.4. DNA Quantification
2.5. Cell Culture
2.6. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR)
2.7. Enzyme-Linked Immunosorbent Assay (ELISA)
2.8. Western Blotting
2.9. Inhibition of MAPK and NF-κB Signaling Pathways
2.10. Knockdown of STING mRNA in OBA-9 Cells Using Small Interfering RNA (siRNA)
2.11. Statistical Analysis
3. Results
3.1. Effect of Pg-OMVs on IL-6 and IL-8 mRNA Levels in OBA-9 Cells
3.2. Pg-OMVs Increased IL-6 and IL-8 mRNA Expression and Protein Production More Than HTPg-OMVs or PgKDP-OMV
3.3. Roles of MAP Kinases in IL-6 and IL-8 Induction by Pg-OMVs
3.4. Involvement of NF-κB p65 in IL-6 and IL-8 Induction by Pg-OMVs
3.5. Effect of STING Knockdown on IL-6 and IL-8 Expression
3.6. Comparison of Pg-OMVs and Fn-OMVs in IL-6 and IL-8 Production by STING Knockdown
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Uemura, Y.; Hiroshima, Y.; Tada, A.; Murakami, K.; Yoshida, K.; Inagaki, Y.; Kuwahara, T.; Murakami, A.; Fujii, H.; Yumoto, H. Porphyromonas gingivalis Outer Membrane Vesicles Stimulate Gingival Epithelial Cells to Induce Pro-Inflammatory Cytokines via the MAPK and STING Pathways. Biomedicines 2022, 10, 2643. https://doi.org/10.3390/biomedicines10102643
Uemura Y, Hiroshima Y, Tada A, Murakami K, Yoshida K, Inagaki Y, Kuwahara T, Murakami A, Fujii H, Yumoto H. Porphyromonas gingivalis Outer Membrane Vesicles Stimulate Gingival Epithelial Cells to Induce Pro-Inflammatory Cytokines via the MAPK and STING Pathways. Biomedicines. 2022; 10(10):2643. https://doi.org/10.3390/biomedicines10102643
Chicago/Turabian StyleUemura, Yuta, Yuka Hiroshima, Ayano Tada, Keiji Murakami, Kaya Yoshida, Yuji Inagaki, Tomomi Kuwahara, Akikazu Murakami, Hideki Fujii, and Hiromichi Yumoto. 2022. "Porphyromonas gingivalis Outer Membrane Vesicles Stimulate Gingival Epithelial Cells to Induce Pro-Inflammatory Cytokines via the MAPK and STING Pathways" Biomedicines 10, no. 10: 2643. https://doi.org/10.3390/biomedicines10102643