Porphyromonas gingivalis stimulates the release of nitric oxide by inducing expression of inducible nitric oxide synthases and inhibiting endothelial nitric oxide synthases

J Periodontal Res. 2010 Jun;45(3):381-8. doi: 10.1111/j.1600-0765.2009.01249.x. Epub 2010 Mar 9.

Abstract

Background and objective: The purpose of this study was to examine the ability of Porphyromonas gingivalis to invade human umbilical vein endothelial cells (HUVECs) and to study the effects of P. gingivalis ATCC 33277 on the production of nitric oxide (NO) and on the expression of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in HUVECs. We attempted to throw light on the pathway of damage to endothelial function induced by P. gingivalis ATCC 33277.

Material and methods: P. gingivalis ATCC 33277 was cultured anaerobically, and HUVECs were treated with P. gingivalis ATCC 33277 at multiplicities of infection of 1:10 or 1:100 for 4, 8, 12 and 24 h. HUVECs were observed using an inverted microscope and transmission electron microscopy. NO production was assayed through measuring the accumulation of nitrite in culture supernatants. Expression of both iNOS and eNOS proteins was investigated through western blotting.

Results: It was found that P. gingivalis ATCC 33277 can adhere to HUVECs by fimbriae, invade into HUVECs and exist in the cytoplasm and vacuoles. P. gingivalis ATCC 33277 can induce iNOS and inhibit eNOS expression, and stimulate the release of NO without any additional stimulant.

Conclusion: Our study provides evidence that P. gingivalis ATCC 33277 can invade HUVECs, and the ability of P. gingivalis ATCC 33277 to promote the production of NO may be important in endothelial dysfunction, suggesting that P. gingivalis ATCC 33277may be one of the pathogens responsible for atherosclerosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Adhesion / physiology
  • Blotting, Western
  • Cell Line
  • Cell Shape
  • Cytoplasm / microbiology
  • Endothelial Cells / enzymology
  • Endothelial Cells / microbiology
  • Endothelium, Vascular / enzymology
  • Endothelium, Vascular / microbiology
  • Enzyme Induction
  • Enzyme Inhibitors / metabolism*
  • Fimbriae, Bacterial / physiology
  • Humans
  • Microscopy, Electron, Transmission
  • Microscopy, Phase-Contrast
  • Nitric Oxide / metabolism*
  • Nitric Oxide Synthase Type II / biosynthesis*
  • Nitric Oxide Synthase Type III / antagonists & inhibitors*
  • Porphyromonas gingivalis / enzymology
  • Porphyromonas gingivalis / metabolism*
  • Vacuoles / microbiology

Substances

  • Enzyme Inhibitors
  • Nitric Oxide
  • NOS2 protein, human
  • NOS3 protein, human
  • Nitric Oxide Synthase Type II
  • Nitric Oxide Synthase Type III