Altered expression of SARS-CoV-2 entry and processing genes by Porphyromonas gingivalis-derived lipopolysaccharide, inflammatory cytokines and prostaglandin E2 in human gingival fibroblasts

Kotaro Sena, Kirara Furue, Fumiaki Setoguchi, Kazuyuki Noguchi, Kotaro Sena, Kirara Furue, Fumiaki Setoguchi, Kazuyuki Noguchi

Abstract

Objective: The aim of this in vitro study was to investigate the expression of SARS-CoV-2 entry and processing genes in human gingival fibroblasts (HGnF) following treatment with Porphyromonas gingivalis-derived lipopolysaccharide (PgLPS) or inflammatory cytokines/mediators.

Design: We assessed the expression of SARS-CoV-2 entry and processing genes; angiotensin-converting enzyme 2 (ACE2), cellular serine proteases transmembrane serine protease 2 (TMPRSS2), Furin, and basigin (BSG) in HGnF by real-time PCR. To further asses the contribution of PgLPS and inflammatory cytokines/mediators to proliferation and SARS-CoV-2 entry and processing gene expression, HGnF were treated with PgLPS, IL1β, TNFα, and PGE2.

Results: The expression for ACE2 in HGnF was significantly elevated after PgLPS or IL1β, TNFα, PGE2 treatment. The expression of TMPRSS2 was increased by PgLPS, IL1β, or PGE2 while BSG was elevated by PgLPS and IL1β. The expression of BSG and FURIN decreased after TNFα treatment.

Conclusion: SARS-CoV-2 entry and processing genes are expressed in human gingival fibroblasts and their expressions are altered by PgLPS, IL1β, TNFα and PGE2 treatment.

Keywords: Angiotensin-converting enzyme 2; Basigin; COVID-19; Furin; Transmembrane serine protease 2.

Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Copyright © 2021 Elsevier Ltd. All rights reserved.

Figures

Fig. 1
Fig. 1
Effect of PgLPS, IL1β, TNFα and PGE2 on HGnF cell proliferation. Cell numbers were analyzed after treatment with (A) PgLPS (0, 0.1, 1, 10 μg/mL), (B) IL1β (0, 0.01, 0.1, 1 ng/mL), (C) TNFα (0, 1, 10, 100 ng/mL) or (D) PGE2 (0, 1, 10, 100, 1000 nM) for 24 h. Data are presented as dot plots. Values are shown as mean ± SD. n = 5. O.D., optical density.
Fig. 2
Fig. 2
Effect of PgLPS, IL1β, TNFα, and PGE2 on expression of ACE2. Expression level of ACE2 mRNA was analyzed after treatment with (A) PgLPS (0, 0.1, 1, 10 μg/mL), (B) IL1β (0, 0.01, 0.1, 1 ng/mL), (C) TNFα (0, 1, 10, 100 ng/mL) or (D) PGE2 (0, 1, 10, 100, 1000 nM) for 24 h. Data are presented as dot plots. Values are shown as mean ± SD. n = 3-8. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Fig. 3
Fig. 3
Effect of PgLPS, IL1β, TNFα, and PGE2 on expression of TMPRSS2. Expression level of TMPRSS2 mRNA was analyzed after treatment with (A) PgLPS (0, 0.1, 1, 10 μg/mL), (B) IL1β (0, 0.01, 0.1, 1 ng/mL), (C) TNFα (0, 1, 10, 100 ng/mL) or (D) PGE2 (0, 1, 10, 100, 1000 nM) for 24 h. Data are presented as dot plots. Values are shown as mean ± SD. n = 3-4. **P < 0.01, ***P < 0.001, ****P < 0.0001.
Fig. 4
Fig. 4
Effect of PgLPS, IL1β, TNFα, and PGE2 on expression of BSG. Expression level of BSG mRNA was analyzed after treatment with (A) PgLPS (0, 0.1, 1, 10 μg/mL), (B) IL1β (0, 0.01, 0.1, 1 ng/mL), (C) TNFα (0, 1, 10, 100 ng/mL) or (D) PGE2 (0, 1, 10, 100, 1000 nM) for 24 h. Data are presented as dot plots. Values are shown as mean ± SD. n = 3. *P < 0.05, **P < 0.01, ****P < 0.0001.
Fig. 5
Fig. 5
Effect of PgLPS, IL1β, TNFα, and PGE2 on expression of FURIN. Expression level of FURIN mRNA was analyzed after treatment with (A) PgLPS (0, 0.1, 1, 10 μg/mL), (B) IL1β (0, 0.01, 0.1, 1 ng/mL), (C) TNFα (0, 1, 10, 100 ng/mL) or (D) PGE2 (0, 1, 10, 100, 1000 nM) for 24 h. Data are presented as dot plots. Values are shown as mean ± SD. n = 3. ****P < 0.0001.

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