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Published on: August 6, 2019
Prevotella intermedia upregulates MMP-1 and MMP-8 expression in human periodontal ligament cells
Su-Min Guan1, Lei Shu, Shan-Min Fu
1Department of Oral Biology, School of Stomatology, The Fourth Military Medical University, 145 Chang Le Xi Road, Xi'an, China. jchsmg@fmmu.edu.cn
Abstract:
Prevotella intermedia, a major periodontal pathogen, plays important roles in the initiation and development of periodontitis by stimulating the release of proinflammatory cytokines, proteinases and matrix metalloproteinases (MMPs). Our previous study demonstrated that P. intermedia induced MMP-9 expression in human periodontal ligament (hPDL) cells. In this study, we examined the effects of P. intermedia on other MMPs' expression. Semi-quantitative reverse transcriptase (RT)-PCR analysis revealed that P. intermedia ATCC 25611 supernatant increased MMP-1 and MMP-8 mRNA expression in a concentration- and time-dependent manner. Enzyme-linked immunosorbent assay and Western blot results confirmed the RT-PCR results at the protein level. Cyclooxygenase inhibitor indomethacin significantly attenuated the upregulatory effects of P. intermedia on MMP-1 and MMP-8 expression. Extracellular signal-related kinase inhibitor PD98059 and c-Jun N-terminal kinase inhibitor SP600125 considerably decreased the upregulated level of MMP-1, whereas p38 inhibitor SB203580 markedly inhibited MMP-8 expression, suggesting that prostaglandin E(2) and mitogen-activated protein kinase signaling pathways are involved in P. intermedia-induced MMP-1 and MMP-8 upregulation. Our results indicate that P. intermedia might contribute to periodontal connective tissue and bone matrix destruction through upregulating MMP production.
Insights
Prevotella intermedia, a periodontal pathogen, upregulates matrix metalloproteinases (MMPs) like MMP-1 and MMP-8. This process involves prostaglandin E2 and MAPK signaling, potentially leading to connective tissue destruction in periodontitis.
Area of Science:
- Oral microbiology
- Molecular biology
- Biochemistry
Background:
- Prevotella intermedia is a key pathogen in periodontitis.
- It stimulates proinflammatory cytokines and matrix metalloproteinases (MMPs).
- Previous work showed P. intermedia induces MMP-9 in human periodontal ligament (hPDL) cells.
Purpose of the Study:
- To investigate the effects of P. intermedia on other MMPs' expression in hPDL cells.
- To elucidate the signaling pathways involved in P. intermedia-induced MMP upregulation.
Main Methods:
- Semi-quantitative reverse transcriptase (RT)-PCR to analyze MMP mRNA expression.
- Enzyme-linked immunosorbent assay (ELISA) and Western blot to confirm protein levels.
- Inhibitors of cyclooxygenase (indomethacin), ERK (PD98059), JNK (SP600125), and p38 (SB203580) were used to study signaling pathways.
Main Results:
- P. intermedia supernatant increased MMP-1 and MMP-8 mRNA and protein expression in a dose- and time-dependent manner.
- Indomethacin significantly reduced P. intermedia-induced MMP-1 and MMP-8 upregulation.
- PD98059 and SP600125 decreased MMP-1 levels, while SB203580 inhibited MMP-8 expression.
Conclusions:
- P. intermedia upregulates MMP-1 and MMP-8 expression in hPDL cells.
- Prostaglandin E2 and mitogen-activated protein kinase (MAPK) signaling pathways mediate this upregulation.
- P. intermedia may contribute to periodontal tissue destruction by increasing MMP production.
