Experimental stress suppresses recruitment of macrophages but enhanced their P. gingivalis LPS-stimulated secretion of nitric oxide.
BACKGROUND
Epidemiological studies have suggested that stress can alter the onset and progression of periodontal disease. However, the mechanisms involved are not clear. The present study was designed to examine whether the functional response of mouse macrophages stimulated by Porphyromonas gingivalis lipopolysaccharide (LPS) is affected by experimental stress, and to investigate the role of corticosterone (CS) in the stress-related effects.


METHODS
Two models of stress were used: emotional (isolation) and physical (cold). We measured thioglycollate-induced macrophage recruitment in vivo, and LPS-induced nitric oxide (NO) secretion by the macrophages in vitro. Two groups of mice were exposed to the stress conditions: isolation or cold. A third group was injected daily with CS, and a fourth group was used as a control (no stress). After 3 days of stress conditions, thioglycollate was injected into the peritoneal cavity. Four days later, peritoneal macrophages were isolated, counted, and cultured. The secretion of NO by the cultured cells was evaluated with and without P. gingivalis LPS stimulation.


RESULTS
The number of cells in the peritoneal lavage of stressed mice was significantly reduced in comparison to macrophages isolated from non-stressed animals. The number of macrophages from CS-treated mice did not differ from controls. NO secretion from unstimulated macrophages did not differ between the stressed and control groups. Stimulation of the macrophages with P. gingivalis LPS significantly enhanced NO secretion by macrophages from the control and stressed animals, but not by the CS-treated group. NO levels secreted by P. gingivalis-stimulated cells from the two stressed groups were significantly higher than the levels secreted by controls, and the isolation group released significantly higher levels than the cold group. Stimulation of the macrophages with P. gingivalis LPS and interferon (IFN)-gamma resulted in enhanced NO secretion in the 4 groups compared to LPS alone, with no significant differences between the groups.


CONCLUSIONS
The results suggest that experimental stress modulates the response of macrophages to inflammatory stimulants, and that CS is not the sole mediator involved. The presence of IFN-gamma in the culture may mask the functional differences induced by stress. The stress-induced upregulation of NO secretion might be involved in the accelerated periodontal destruction in stressed subjects.
Shapira
L
L
Department of Periodontology, Hebrew University-Hasassah School of Dental Medicine, Jerusalem, Israel. shapiral@cc.huji.ac.il
Frolov
I
I
Halabi
A
A
Ben-Nathan
D
D
D000704:Analysis of Variance; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002452:Cell Count; D002469:Cell Separation; D002478:Cells, Cultured; D003345:Corticosterone / Q000494:pharmacology; D005260:Female; D016166:Free Radical Scavengers / Q000276:immunology / Q000378:metabolism*; D007371:Interferon-gamma / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology*; D008262:Macrophage Activation / Q000187:drug effects / Q000276:immunology*; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000276:immunology / Q000378:metabolism*; D016966:Porphyromonas gingivalis / Q000276:immunology*; D013312:Stress, Physiological / Q000276:immunology*; D013315:Stress, Psychological / Q000276:immunology; D013864:Thioglycolates / Q000494:pharmacology; D015854:Up-Regulation
D000893:Anti-Inflammatory Agents; D016166:Free Radical Scavengers; D008070:Lipopolysaccharides; D013864:Thioglycolates; D009569:Nitric Oxide; D007371:Interferon-gamma; D003345:Corticosterone