Abstract
Objective
Porphyromonas gingivalis is a major periodontopathogen that plays a role in the pathogenesis of periodontal disease. In this study, we investigated the effect of 18alpha-glycyrrhetinic acid (18α-GA), a natural triterpenoid compound derived from licorice root extract, on P. gingivalis lipopolysaccharide (LPS)-induced vascular permeability, which is a hallmark of inflammatory diseases such as periodontitis.
Methods
The inhibitory effects of 18α-GA on endothelial permeability were determined by measuring in vivo and in vitro endothelial permeability. Endothelial cells were pretreated with 18α-GA before exposure to P. gingivalis LPS, and total RNA or proteins were extracted and analyzed by reverse transcription polymerase chain reaction or western blotting.
Results
Porphyromonas gingivalis LPS-induced endothelial permeability was significantly inhibited by 18α-GA both in vivo and in vitro. 18α-GA reduces P. gingivalis LPS-induced gap formation of endothelial cells. Importantly, 18α-GA modulated the expression and secretion of interleukin-8 (IL-8), a key inducer of vascular permeability, by downregulating nuclear factor-κB (NF-κB). 18α-GA suppressed P. gingivalis LPS-stimulated inhibitor of kappa B (IκB) kinase activation, IκBα phosphorylation, and nuclear translocation of NF-κB.
Conclusions
Overall, these findings suggest that 18α-GA significantly reduces P. gingivalis LPS-induced vascular permeability by repressing NF-κB-dependent endothelial IL-8 production, suggesting its therapeutic potential in P. gingivalis-related vascular diseases.
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Acknowledgments
This research was supported by College of Pharmacy-specialized Research Fund (from institute for new drug development) of Keimyung University in 2011 (to C.-H. Jeong) and by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2010-0003750) (to M.-K. Bae).
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Responsible Editor: Michael Parnham.
S.-R. Kim and H.-J. Jeon are co-first authors.
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11_2012_560_MOESM1_ESM.ppt
Supplemental Figure 1. Immunocytochemical views of PECAM-1 with a low magnification. HUVECs incubated with P. gingivalis LPS (10 μg/ml) alone or in combination with 18α-GA (100 nM) for 12 h were immunostained for PECAM-1, and then photographed under fluorescence microscope (10X). The arrows indicate the position of the paracellular gaps
Supplemental Figure 2. Effect of recombinant IL-8 protein on paracellular gap formation in endothelial cells. HUVECs were treated with P. gingivalis LPS (10 μg/ml) or IL-8 (10 ng/ml) in the presence or absence of anti-IL-8 (1 μg/ml) and immunostained with anti-PECAM-1
Supplemental Figure 3. The schematic diagram of our proposed mechanism of the inhibitory effect of 18α-GA on the P. gingivalis LPS-induced endothelial permeability. (PPT 1894 kb)
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Kim, SR., Jeon, HJ., Park, HJ. et al. Glycyrrhetinic acid inhibits Porphyromonas gingivalis lipopolysaccharide-induced vascular permeability via the suppression of interleukin-8. Inflamm. Res. 62, 145–154 (2013). https://doi.org/10.1007/s00011-012-0560-5
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DOI: https://doi.org/10.1007/s00011-012-0560-5