N-acetylcysteine and the human serum components that inhibit bacterial invasion of gingival epithelial cells prevent experimental periodontitis in mice
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose: We previously reported that human serum significantly reduces the invasion ofvarious oral bacterial species into gingival epithelial cells in vitro. The aims of the presentstudy were to characterize the serum component(s) responsible for the inhibition of bacterialinvasion of epithelial cells and to examine their effect on periodontitis induced in mice. Methods: Immortalized human gingival epithelial (HOK-16B) cells were infected with various5- (and 6-) carboxy-fluorescein diacetate succinimidyl ester-labeled oral bacteria, includingFusobacterium nucleatum, Provetella intermedia, Porphyromonas gingivalis, andTreponiema denticola, in the absence or presence of three major serum components (humanserum albumin [HSA], pooled human IgG [phIgG] and α1-antitrypsin). Bacterial adhesionand invasion were determined by flow cytometry. The levels of intracellular reactiveoxygen species (ROS) and activation of small GTPases were examined. Experimental periodontitiswas induced by oral inoculation of P. gingivalis and T. denticola in Balb/c mice. Results: HSA and phIgG, but not α1-antitrypsin, efficiently inhibited the invasion of variousoral bacterial species into HOK-16B cells. HSA but not phIgG decreased the adhesion ofF. nucleatum onto host cells and the levels of intracellular ROS in HOK-16B cells. N-acetylcysteine(NAC), a ROS scavenger, decreased both the levels of intracellular ROS and invasionof F. nucleatum into HOK-16B cells, confirming the role of ROS in bacterial invasion. Infection with F. nucleatum activated Rac1, a regulator of actin cytoskeleton dynamics. Not only HSA and NAC but also phIgG decreased the F. nucleatum-induced activation ofRac1. Furthermore, both HSA plus phIgG and NAC significantly reduced the alveolar boneloss in the experimental periodontitis induced by P. gingivalis and T. denticola in mice. Conclusions: NAC and the serum components HSA and phIgG, which inhibit bacterial invasionof oral epithelial cells in vitro, can successfully prevent experimental periodontitis.
Abstract
Purpose: We previously reported that human serum significantly reduces the invasion ofvarious oral bacterial species into gingival epithelial cells in vitro. The aims of the presentstudy were to characterize the serum component(s) responsible for the inhibition of bacterialinvasion of epithelial cells and to examine their effect on periodontitis induced in mice. Methods: Immortalized human gingival epithelial (HOK-16B) cells were infected with various5- (and 6-) carboxy-fluorescein diacetate succinimidyl ester-labeled oral bacteria, includingFusobacterium nucleatum, Provetella intermedia, Porphyromonas gingivalis, andTreponiema denticola, in the absence or presence of three major serum components (humanserum albumin [HSA], pooled human IgG [phIgG] and α1-antitrypsin). Bacterial adhesionand invasion were determined by flow cytometry. The levels of intracellular reactiveoxygen species (ROS) and activation of small GTPases were examined. Experimental periodontitiswas induced by oral inoculation of P. gingivalis and T. denticola in Balb/c mice. Results: HSA and phIgG, but not α1-antitrypsin, efficiently inhibited the invasion of variousoral bacterial species into HOK-16B cells. HSA but not phIgG decreased the adhesion ofF. nucleatum onto host cells and the levels of intracellular ROS in HOK-16B cells. N-acetylcysteine(NAC), a ROS scavenger, decreased both the levels of intracellular ROS and invasionof F. nucleatum into HOK-16B cells, confirming the role of ROS in bacterial invasion. Infection with F. nucleatum activated Rac1, a regulator of actin cytoskeleton dynamics. Not only HSA and NAC but also phIgG decreased the F. nucleatum-induced activation ofRac1. Furthermore, both HSA plus phIgG and NAC significantly reduced the alveolar boneloss in the experimental periodontitis induced by P. gingivalis and T. denticola in mice. Conclusions: NAC and the serum components HSA and phIgG, which inhibit bacterial invasionof oral epithelial cells in vitro, can successfully prevent experimental periodontitis.
참고문헌 (33)
- Dale BA, "Periodontal epithelium: a newly recognized role in health and disease", Periodontol 2000, 30, pp.70~78 (2002)
- Choi YS, "Increased bacterial invasion and differential expression of tight-junction proteins, growth factors, and growth factor receptors in periodontal lesions", J Periodontol, 85, pp.e313~e322 (2014)
- Dixon DR, "Modulation of the innate immune response within the periodontium", Periodontol 2000, 35, pp.53~74 (2004)
- Ji S1, "Bacterial invasion and persistence: critical events in the pathogenesis of periodontitis?", J Periodontal Res (2014)
- Lamont RJ, "In or out: the invasiveness of oral bacteria", Periodontol 2000, 30, pp.61~69 (2002)
- Suk Ji, "Intracellular Degradation of Fusobacterium nucleatum in Human Gingival Epithelial Cells", Molecules and Cells, 30(6), pp.519~526 (2010), 한국분자세포생물학회
- Kim Y, "Toll-like receptor 9 mediates oral bacteria-induced IL-8 expression in gingival epithelial cells", Immunol Cell Biol, 90, pp.655~663 (2012)
- Carneiro LG, "Proteome data set of human gingival crevicular fluid from healthy periodontium sites by multidimensional protein separation and mass spectrometry", J Periodontal Res, 47, pp.248~262 (2012)
- Guentsch A, "Influence of serum on interaction of Porphyromonas gingivalis ATCC 33277 and Aggregatibacter actinomycetemcomitans Y4 with an epithelial cell line", J Periodontal Res, 45, pp.229~238 (2010)
- Baek KJ, "Gingival fibroblasts from periodontitis patients exhibit inflammatory characteristics in vitro", ARCHIVES OF ORAL BIOLOGY, 58, pp.1282~1292 (2013), PERGAMON-ELSEVIER SCIENCE LTD
- Baker PJ, "Oral infection with Porphyromonas gingivalis and induced alveolar bone loss in immunocompetent and severe combined immunodeficient mice", Arch Oral Biol, 39, pp.1035~1040 (1994)
- Martosella J, "Multi-component immunoaffinity subtraction and reversed-phase chromatography of human serum", Methods Mol Biol, 425, pp.27~39 (2008)
- Socransky SS, "Microbial complexes in subgingival plaque", J Clin Periodontol, 25, pp.134~144 (1998)
- Taverna M, "Specific antioxidant properties of human serum albumin", Ann Intensive Care, 3, pp.4~ (2013)
- Brown DI, "Nox proteins in signal transduction", Free Radic Biol Med, 47, pp.1239~1253 (2009)
- Taulet N, "Reactive oxygen species regulate protrusion efficiency by controlling actin dynamics", PLoS One, 7, pp.e41342~ (2012)
- Van Aelst L, "Rho GTPases and signaling networks", Genes Dev, 11, pp.2295~2322 (1997)
- Katsikogianni M, "Concise review of mechanisms of bacterial adhesion to biomaterials and of techniques used in estimating bacteria-material interactions", Eur Cell Mater, 8, pp.37~57 (2004)
- Brokke P, "Adherence of coagulase-negative staphylococci onto polyethylene catheters in vitro and in vivo: a study on the influence of various plasma proteins", J Biomater Appl, 5, pp.204~226 (1991)
- Boks NP, "Forces involved in bacterial adhesion to hydrophilic and hydrophobic surfaces", Microbiology, 154(Pt 10), pp.3122~3133 (2008)
- Su H, "Chlamydia trachomatis-host cell interactions: role of the chlamydial major outer mem-brane protein as an adhesin", Infect Immun, 58, pp.1017~1025 (1990)
- Burnham CA, "Rac1, RhoA, and Cdc42 participate in HeLa cell invasion by group B streptococcus", FEMS Microbiol Lett, 272, pp.8~14 (2007)
- Criss AK, "The GTPase Rac1 selectively regulates Salmonella invasion at the apical plasma membrane of polarized epithelial cells", J Cell Sci, 114(Pt 7), pp.1331~1341 (2001)
- Krause-Gruszczynska M, "Role of the small Rho GTPases Rac1 and Cdc42 in host cell invasion of Campylobacter jejuni", Cell Microbiol, 9, pp.2431~2444 (2007)
- Ridley AJ, "The small GTP-binding protein rac regulates growth factor-induced membrane ruffling", Cell, 70, pp.401~410 (1992)
- Kurokawa K, "Coactivation of Rac1 and Cdc42 at lamellipodia and membrane ruffles induced by epidermal growth factor", Mol Biol Cell, 15, pp.1003~1010 (2004)
- Njoroge T, "A role for fimbriae in Porphyromonas gingivalis invasion of oral epithelial cells", Infect Immun, 65, pp.1980~1984 (1997)
- Ikegami A, "Complementation of the fadA mutation in Fusobacterium nucleatum demonstrates that the surface-exposed adhesin promotes cellular invasion and placental colonization", Infect Immun, 77, pp.3075~3079 (2009)
- Shin J, "Antibody and T cell responses to Fusobacterium nucleatum and Treponema denticola in health and chronic periodontitis", PLoS One, 8, pp.e53703~ (2013)
- Furuichi Y, "Periodontal status and serum antibody titers for Porphyromonas gingivalis fimbriae in a rural population in Japan", J Clin Periodontol, 28, pp.264~269 (2001)
- Yilmaz O, "Involvement of integrins in fimbriae-mediated binding and invasion by Porphyromonas gingivalis", Cell Microbiol, 4, pp.305~314 (2002)
- Busscher HJ, "Specific and non-specific interactions in bacterial adhesion to solid substrata", FEMS Microbiol Lett, 46, pp.165~173 (1987)
- Toker H, "N-acetylcysteine, a thiol antioxidant, decreases alveolar bone loss in experimental periodontitis in rats", J Periodontol, 80, pp.672~678 (2009)