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The effect of bile acids on Porphyromonas gingivalis lipopolysaccharide-induced inflammatory respo

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¹ÚÁö¾Æ ( Park Ji-A ) - Seoul National University School of Dentistry Department of Preventive and Social Dentistry

Abstract


Objectives: In this study, I aimed to evaluate the inhibitory effect of bile acids on the inflammatory response and osteoclastogenesis in RAW 264.7 cells activated through lipopolysaccharide (LPS) and receptor activator of nuclear factor-kB ligand (RANKL) of the periodontal pathogen Porphyromonas gingivalis.

Methods: Myelomonocytic RAW 264.7 cells were activated through P. gingivalis LPS to induce inflammatory response, and were treated with three bile acids, including taurodeoxycholate, taurocholate, and glycocholate at different concentrations. The cytotoxicity of bile acids was assessed through the MTT assay. To evaluate the inhibitory effect of bile acids on inflammatory response, the induction levels of the pro-inflammatory cytokines, such as interleukin (IL)-6 and tumor necrosis factor (TNF)-a were measured using ELISA 12 h after the treatment. Additionally, after activating the cells with RANKL to promote osteoclastogenesis, we examined whether bile acids suppressed osteoclast differentiation using the tartrate-resistant acid phosphatase staining.

Results: In the cell viability test, taurodeoxycholate and taurocholate did not exhibit any cytotoxic effect on RAW 264.7 cells at concentrations equal to or less than 200 mM, and glycocholate was non-cytotoxic until the maximal concentration (4,000 mM). All the three bile acids exhibited an inhibitory effect on inflammatory response, as the production levels of pro-inflammatory cytokines, including IL-6 and TNF-a, decreased with an increase in the concentration of the three bile acids in a dose-dependent manner. The expression of IL-6 reduced remarkably upon treatment with taurodeoxycholate and glycocholate (P<0.001), while the expression of TNF-a decreased slightly upon treatment with glycocholate (P<0.05). Moreover, only glycocholate at a concentration of 1,000 mM suppressed osteoclast differentiation of RAW 264.7 cells (P<0.001), while taurodeoxycholate and taurocholate did not exhibit an inhibitory effect on osteoclastogenesis.

Conclusions: Here, I showed that all the three bile acids (taurodeoxycholate, taurocholate, and glycocholate) inhibited P. gingivalis LPS-induced inflammatory response, and glycocholate partially suppressed RANKL-mediated osteoclastogenesis in RAW 264.7 cells.

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Bile acids; Glycocholic acid; Inflammation; Periodontitis; Porphyromonas gingivalis; Taurodeoxycholic acid

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