Abstract
Nitro-conjugated linoleic acid (NO2-CLA) has been observed to manifest salutary signaling responses, including anti-inflammatory and antioxidant properties. Here, the authors have explored the influence and underlying mechanisms of NO2-CLA on the proinflammatory reaction of murine macrophages that were challenged with lipopolysaccharide (LPS) derived from Prevotella intermedia, a putative periodontopathic bacterium. Treatment of LPS-activated RAW264.7 cells with NO2-CLA notably dampened the secretion of iNOS-derived NO, IL-1β and IL-6 as well as their gene expressions and significantly enhanced the markers for M2 macrophage polarization. NO2-CLA promoted the HO-1 expression in cells challenged with LPS, and tin protoporphyrin IX, an HO-1 inhibitor, significantly reversed the NO2-CLA-mediated attenuation of NO secretion, but not IL-1β or IL-6. We found that cells treated with NO2-CLA significantly increased mRNA expression of PPAR-γ compared to control cells, and NO2-CLA significantly reverted the decrease in PPAR-γ mRNA caused by LPS. Nonetheless, antagonists to PPAR-γ were unable to reverse the NO2-CLA-mediated suppression of inflammatory mediators. In addition, NO2-CLA did not alter the p38 and JNK activation elicited by LPS. Both NF-κB reporter activity and IκB-α degradation caused by LPS were notably diminished by NO2-CLA. NO2-CLA was observed to interrupt the nuclear translocation and DNA binding of p50 subunits caused by LPS with no obvious alterations in p65 subunits. Further, NO2-CLA attenuated the phosphorylation of STAT1/3 elicited in response to LPS. We propose that NO2-CLA could be considered as a possible strategy for the therapy of periodontal disease, although additional researches are certainly required to confirm this.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The obtained data analyzed during the current study are available from the corresponding author on reasonable request.
References
Awwad K, Steinbrink SD, Frömel T, Lill N, Isaak J, Häfner AK et al (2014) Electrophilic fatty acid species inhibit 5-lipoxygenase and attenuate sepsis-induced pulmonary inflammation. Antioxid Redox Signal 20:2667–2680. https://doi.org/10.1089/ars.2013.5473
Baeuerle PA, Henkel T (1994) Function and activation of NF-κB in the immune system. Annu Rev Immunol 12:141–179. https://doi.org/10.1146/annurev.iy.12.040194.001041
Baker PR, Lin Y, Schopfer FJ, Woodcock SR, Groeger AL, Batthyany C et al (2005) Fatty acid transduction of nitric oxide signaling: multiple nitrated unsaturated fatty acid derivatives exist in human blood and urine and serve as endogenous peroxisome proliferator-activated receptor ligands. J Biol Chem 280:42464–42475. https://doi.org/10.1074/jbc.M504212200
Beem JE, Nesbitt WE, Leung KP (1998) Identification of hemolytic activity in Prevotella intermedia. Oral Microbiol Immunol 13:97–105. https://doi.org/10.1111/j.1399-302x.1998.tb00719.x
Biswas SK, Mantovani A (2012) Orchestration of metabolism by macrophages. Cell Metab 5:432–437. https://doi.org/10.1016/j.cmet.2011.11.013
Bonacci G, Baker PR, Salvatore SR, Shores D, Khoo NK, Koenitzer JR et al (2012) Conjugated linoleic acid is a preferential substrate for fatty acid nitration. J Biol Chem 287:44071–44082. https://doi.org/10.1074/jbc.M112.401356.
Borniquel S, Jansson EA, Cole MP, Freeman BA, Lundberg JO (2010) Nitrated oleic acid up-regulates PPARgamma and attenuates experimental inflammatory bowel disease. Free Radic Biol Med 48:499–505. https://doi.org/10.1016/j.freeradbiomed.2009.11.014
Bouhlel MA, Derudas B, Rigamonti E, Dièvart R, Brozek J, Haulon S et al (2007) PPARγ activation primes human monocytes into alternative M2 macrophages with anti-inflammatory properties. Cell Metab 6:137–143. https://doi.org/10.1016/j.cmet.2007.06.010
Braga RR, Carvalho MA, Bruña-Romero O, Teixeira RE, Costa JE, Mendes EN et al (2010) Quantification of five putative periodontal pathogens in female patients with and without chronic periodontitis by real-time polymerase chain reaction. Anaerobe 16:234–239. https://doi.org/10.1016/j.anaerobe.2010.02.007
Brook I, Frazier EH (2003) Immune response to Fusobacterium nucleatum and Prevotella intermedia in the sputum of patients with acute exacerbation of chronic bronchitis. Chest 124:832–833. https://doi.org/10.1378/chest.124.3.832
Cekici A, Kantarci A, Hasturk H, Van Dyke TE (2014) Inflammatory and immune pathways in the pathogenesis of periodontal disease. Periodontol 64:57–80. https://doi.org/10.1111/prd.12002
Choi EY, ** JY, Lee JY, Choi JI, Choi IS, Kim SJ (2011) Melatonin inhibits Prevotella intermedia lipopolysaccharide-induced production of nitric oxide and interleukin-6 in murine macrophages by suppressing NF-κB and STAT1 activity. J Pineal Res 50:197–206. https://doi.org/10.1111/j.1600-079X.2010.00829.x
Choi EY, ** JY, Choi JI, Choi IS, Kim SJ (2014) DHA suppresses Prevotella intermedia lipopolysaccharide-induced production of proinflammatory mediators in murine macrophages. Br J Nutr 111:1221–1230. https://doi.org/10.1017/S0007114513003681
Choi EY, Choe SH, Hyeon JY, Choi JI, Choi IS, Kim SJ (2015) Effect of caffeic acid phenethyl ester on Prevotella intermedia lipopolysaccharide-induced production of proinflammatory mediators in murine macrophages. J Periodontal Res 50:737–747. https://doi.org/10.1111/jre.12260
Chung CP, Nisengard RJ, Slots J, Genco RJ (1983) Bacterial IgG and IgM antibody titers in acute necrotizing ulcerative gingivitis. J Periodontol 54:557–562. https://doi.org/10.1902/jop.1983.54.9.557
Delmastro-Greenwood M, Hughan KS, Vitturi DA, Salvatore SR, Grimes G, Potti G et al (2015) Nitrite and nitrate-dependent generation of anti-inflammatory fatty acid nitroalkenes. Free Radic Biol Med 89:333–341. https://doi.org/10.1016/j.freeradbiomed.2015.07.149
Gao JJ, Filla MB, Fultz MJ, Vogel SN, Russell SW, Murphy WJ et al (1998) Autocrine/paracrine IFN-αβ mediates the lipopolysaccharide-induced activation of transcription factor Stat1α in mouse macrophages: pivotal role of Stat1α in induction of the inducible nitric oxide synthase gene. J Immunol 161:4803–4810
Ghosh S, May MJ, Kopp EB (1998) NF-κB and Rel proteins: evolutionarily conserved mediators of immune responses. Annu Rev Immunol 16:225–260. https://doi.org/10.1146/annurev.immunol.16.1.225
Guan SM, Shu L, Fu SM, Liu B, Xu XL, Wu JZ (2009) Prevotella intermedia upregulates MMP-1 and MMP-8 expression in human periodontal ligament cells. FEMS Microbiol Lett 299:214–222. https://doi.org/10.1111/j.1574-6968.2009.01748.x
Hamada S, Takada H, Ogawa T, Fujiwara T, Mihara J (1990) Lipopolysaccharides of oral anaerobes associated with chronic inflammation: chemical and immunomodulating properties. Int Rev Immunol 6:247–261. https://doi.org/10.3109/08830189009056635
Hayden MS, Ghosh S (2004) Signaling to NF-κB. Genes Dev 18:2195–2224. https://doi.org/10.1101/gad.1228704
Irwin CR, Myrillas TT (1998) The role of IL-6 in the pathogenesis of periodontal disease. Oral Dis 4:43–47. https://doi.org/10.1111/j.1601-0825.1998.tb00255.x
Ivashkiv LB (2013) Epigenetic regulation of macrophage polarization and function. Trends Immunol 34:216–223. https://doi.org/10.1016/j.it.2012.11.001
Jacinto RC, Gomes BP, Ferraz CC, Zaia AA, Filho FJ (2003) Microbiological analysis of infected root canals from symptomatic and asymptomatic teeth with periapical periodontitis and the antimicrobial susceptibility of some isolated anaerobic bacteria. Oral Microbiol Immunol 18:285–292. https://doi.org/10.1034/j.1399-302x.2003.00078.x
Karin M, Ben-Neriah Y (2000) Phosphorylation meets ubiquitination: the control of NF-κB activity. Annu Rev Immunol 18:621–663. https://doi.org/10.1146/annurev.immunol.18.1.621
Khan SA, Kong EF, Meiller TF, Jabra-Rizk MA (2015) Periodontal Diseases: Bug Induced, Host Promoted. PLoS Pathog 11:e1004952. https://doi.org/10.1371/journal.ppat.1004952.
Kim SJ, Ha MS, Choi EY, Choi JI, Choi IS (2004) Prevotella intermedia lipopolysaccharide stimulates release of nitric oxide by inducing expression of inducible nitric oxide synthase. J Periodontal Res 39:424–431. https://doi.org/10.1111/j.1600-0765.2004.00757.x
Kim SJ, Choi EY, Kim EG, Shin SH, Lee JY, Choi JI et al (2007) Prevotella intermedia lipopolysaccharide stimulates release of tumor necrosis factor-alpha through mitogen-activated protein kinase signaling pathways in monocyte-derived macrophages. FEMS Immunol Med Microbiol 51:407–413. https://doi.org/10.1111/j.1574-695X.2007.00318.x
Kirikae T, Nitta T, Kirikae F, Suda Y, Kusumoto S, Qureshi N et al (1999) Lipopolysaccharides (LPS) of oral black-pigmented bacteria induce tumor necrosis factor production by LPS-refractory C3H/HeJ macrophages in a way different from that of Salmonella LPS. Infect Immun 67:1736–1742. https://doi.org/10.1128/IAI.67.4.1736-1742.1999
Kobayashi K, Takahashi N, Jimi E, Udagawa N, Takami M, Kotake S et al (2000) Tumor necrosis factor alpha stimulates osteoclast differentiation by a mechanism independent of the ODF/RANKL-RANK interaction. J Exp Med 191:275–286. https://doi.org/10.1084/jem.191.2.275
Kornman KS, Loesch WJ (1980) The subgingival microbial flora during pregnancy. J Periodontal Res 15:111–122. https://doi.org/10.1111/j.1600-0765.1980.tb00265.x
Kühn B, Brat C, Fettel J, Hellmuth N, Maucher IV, Bulut U et al (2018) Anti-inflammatory nitro-fatty acids suppress tumor growth by triggering mitochondrial dysfunction and activation of the intrinsic apoptotic pathway in colorectal cancer cells. Biochem Pharmacol 155:48–60. https://doi.org/10.1016/j.bcp.2018.06.014
Lam RS, O’Brien-Simpson NM, Lenzo JC, Holden JA, Brammar GC, Walsh KA et al (2014) Macrophage depletion abates Porphyromonas gingivalis-induced alveolar bone resorption in mice. J Immunol 193:2349–2362. https://doi.org/10.4049/jimmunol.1400853
Leonard WJ, O’Shea JJ (1998) Jaks and STATs: biological implications. Annu Rev Immunol 16:293–322. https://doi.org/10.1146/annurev.immunol.16.1.293
Li Q, Verma IM (2002) NF-κB regulation in the immune system. Nat Rev Immunol 2:725–734. https://doi.org/10.1038/nri910
Liu XH, Kirschenbaum A, Yao S, Levine AC (2005) Cross-talk between the interleukin-6 and prostaglandin E2 signaling systems results in enhancement of osteoclastogenesis through effects on the osteoprotegerin/receptor activator of NF-κB (RANK) ligand/RANK system. Endocrinology 146:1991–1998. https://doi.org/10.1210/en.2004-1167
Matejka M, Partyka L, Ulm C, Solar P, Sinzinger H (1998) Nitric oxide synthesis is increased in periodontal disease. J Periodontal Res 33:517–518. https://doi.org/10.1111/j.1600-0765.1998.tb02352.x
Mogi M, Otogoto J, Ota N, Inagaki H, Minami M, Kojima K (1999) Interleukin 1β, interleukin 6, β2-microglobulin, and transforming growth factor-α in gingival crevicular fluid from human periodontal disease. Arch Oral Biol 44:535–539. https://doi.org/10.1016/s0003-9969(99)00020-5
Morrison DC, Ryan JL (1987) Endotoxins and disease mechanisms. Annu Rev Med 38:417–432. https://doi.org/10.1146/annurev.me.38.020187.002221
Morse D, Choi AM (2002) Heme oxygenase-1: the “emerging molecule” has arrived. Am J Respir Cell Mol Biol 27:8–16. https://doi.org/10.1165/ajrcmb.27.1.4862
Morse D, Pischke SE, Zhou Z, Davis RJ, Flavell RA, Loop T et al (2003) Suppression of inflammatory cytokine production by carbon monoxide involves the JNK pathway and AP-1. J Biol Chem 278:36993–36998. https://doi.org/10.1074/jbc.M302942200
Mosser DM, Edwards JP (2008) Exploring the full spectrum of macrophage activation. Nat Rev Immunol 8:958–969. https://doi.org/10.1038/nri2448
Otterbein LE, Bach FH, Alam J, Soares M, Tao LuH, Wysk M et al (2000) Carbon monoxide has anti-inflammatory effects involving the mitogen-activated protein kinase pathway. Nat Med 6:422–428. https://doi.org/10.1038/74680
Otterbein LE, Soares MP, Yamashita K, Bach FH (2003) Heme oxygenase-1: unleashing the protective properties of heme. Trends Immunol 24:449–455. https://doi.org/10.1016/s1471-4906(03)00181-9
Pelt P, Zimmermann B, Ulbrich N, Bernimoulin JP (2002) Effects of lipopolysaccharide extracted from Prevotella intermedia on bone formation and on the release of osteolytic mediators by fetal mouse osteoblasts in vitro. Arch Oral Biol 47:859–866. https://doi.org/10.1016/s0003-9969(02)00150-4
Pfitzner E, Kliem S, Baus D, Litterst CM (2004) The role of STATs in inflammation and inflammatory diseases. Curr Pharm Des 10:2839–2850. https://doi.org/10.2174/1381612043383638
Poss KD, Tonegawa S (1997) Reduced stress defense in heme oxygenase 1-deficient cells. Proc Natl Acad Sci USA 94:10925–10930. https://doi.org/10.1073/pnas.94.20.10925
Preshaw PM (2008) Host Response Modulation in Periodontics Periodontol 2000(48):92–110. https://doi.org/10.1111/j.1600-0757.2008.00252.x
Pucar A, Milasin J, Lekovic V, Vukadinovic M, Ristic M, Putnik S et al (2007) Correlation between atherosclerosis and periodontal putative pathogenic bacterial infections in coronary and internal mammary arteries. J Periodontol 78:677–682. https://doi.org/10.1902/jop.2007.060062
Reddy MS, Geurs NC, Gunsolley JC (2003) Periodontal host modulation with antiproteinase, anti-inflammatory, and bone-sparing agents. A systematic review. Ann Periodontol 8:12–37. https://doi.org/10.1902/annals.2003.8.1.12.
Reddy AT, Lakshmi SP, Dornadula S, Pinni S, Rampa DR, Reddy RC (2013) The nitrated fatty acid 10-nitro-oleate attenuates allergic airway disease. J Immunol 191:2053–2063. https://doi.org/10.4049/jimmunol.1300730.
Ricote M, Glass CK (2007) PPARs and molecular mechanisms of transrepression. Biochim Biophys Acta 1771:926–935. https://doi.org/10.1016/j.bbalip.2007.02.013
Rossomando EF, Kennedy JE, Hadjimichael J (1990) Tumour necrosis factor alpha in gingival crevicular fluid as a possible indicator of periodontal disease in humans. Arch Oral Biol 35:431–434. https://doi.org/10.1016/0003-9969(90)90205-o
Rudolph V, Rudolph TK, Schopfer FJ, Bonacci G, Woodcock SR, Cole MP et al (2010a) Endogenous generation and protective effects of nitro-fatty acids in a murine model of focal cardiac ischaemia and reperfusion. Cardiovasc Res 85:155–166. https://doi.org/10.1093/cvr/cvp275
Rudolph TK, Rudolph V, Edreira MM, Cole MP, Bonacci G, Schopfer FJ et al (2010b) Nitro-fatty acids reduce atherosclerosis in apolipoprotein E-deficient mice. Arterioscler Thromb Vasc Biol 30:938–945. https://doi.org/10.1161/ATVBAHA.109.201582
Ryter SW, Alam J, Choi AM (2006) Heme oxygenase-1/carbon monoxide: from basic science to therapeutic applications. Physiol Rev 86:583–650. https://doi.org/10.1152/physrev.00011
Saijo K, Crotti A, Glass CK (2013) Regulation of microglia activation and deactivation by nuclear receptors. Glia 61:104–111. https://doi.org/10.1002/glia.22423
Salvatore SR, Vitturi DA, Baker PR, Bonacci G, Koenitzer JR, Woodcock SR et al (2013) Characterization and quantification of endogenous fatty acid nitroalkene metabolites in human urine. J Lipid Res 54:1998–2009. https://doi.org/10.1194/jlr.M037804
Samavati L, Rastogi R, Du W, Hüttemann M, Fite A, Franchi L (2009) STAT3 tyrosine phosphorylation is critical for interleukin 1 beta and interleukin-6 production in response to lipopolysaccharide and live bacteria. Mol Immunol 46:1867–1877. https://doi.org/10.1016/j.molimm.2009.02.018
Sarady-Andrews JK, Liu F, Gallo D, Nakao A, Overhaus M, Ollinger R et al (2005) Biliverdin administration protects against endotoxin-induced acute lung injury in rats. Am J Physiol Lung Cell Mol Physiol 289:L1131-1137. https://doi.org/10.1152/ajplung.00458.2004
Schindler C, Levy DE, Decker T (2007) JAK-STAT signaling: from interferons to cytokines. J Biol Chem 282:20059–20063. https://doi.org/10.1074/jbc.R700016200
Schopfer FJ, Lin Y, Baker PR, Cui T, Garcia-Barrio M, Zhang J et al (2005) Nitrolinoleic acid: an endogenous peroxisome proliferator-activated receptor gamma ligand. Proc Natl Acad Sci USA 102:2340–2345. https://doi.org/10.1073/pnas.0408384102
Schopfer FJ, Cipollina C, Freeman BA (2011) Formation and signaling actions of electrophilic lipids. Chem Rev 111:5997–6021. https://doi.org/10.1021/cr200131e
Schopfer FJ, Vitturi DA, Jorkasky DK, Freeman BA (2018) Nitro-fatty acids: new drug candidates for chronic inflammatory and fibrotic diseases. Nitric Oxide 79:31–37. https://doi.org/10.1016/j.niox.2018.06.006
Seymour GJ, Ford PJ, Cullinan MP, Leishman S, Yamazaki K (2007) Relationship between periodontal infections and systemic disease. Clin Microbiol Infect 13(Suppl 4):3–10. https://doi.org/10.1111/j.1469-0691.2007.01798.x
Shenker BJ, Vitale L, Slots J (1991) Immunosuppressive effects of Prevotella intermedia on in vitro human lymphocyte activation. Infect Immun 59:4583–4589. https://doi.org/10.1128/iai.59.12.4583-4589
Stout RD, Jiang C, Matta B, Tietzel I, Watkins SK, Suttles J (2005) Macrophages sequentially change their functional phenotype in response to changes in microenvironmental influences. J Immunol 175:342–349. https://doi.org/10.4049/jimmunol.175.1.342
Tatakis DN (1993) Interleukin-1 and bone metabolism: a review. J Periodontol 64(5 Suppl):416–431
Trostchansky A, Bonilla L, González-Perilli L, Rubbo H (2013) Nitro-fatty acids: formation, redox signaling, and therapeutic potential. Antioxid Redox Signal 19:1257–1265. https://doi.org/10.1089/ars.2012.5023
Ulrich M, Beer I, Braitmaier P, Dierkes M, Kummer F, Krismer B et al (2010) Relative contribution of Prevotella intermedia and Pseudomonas aeruginosa to lung pathology in airways of patients with cystic fibrosis. Thorax 65:978–984. https://doi.org/10.1136/thx.2010.137745
Villacorta L, Gao Z, Schopfer FJ, Freeman BA, Chen YE (2016) Nitro-fatty acids in cardiovascular regulation and diseases: characteristics and molecular mechanisms. Front Biosci (landmark Ed) 21:873–889. https://doi.org/10.2741/4425
Villacorta L, Minarrieta L, Salvatore SR, Khoo NK, Rom O, Gao Z et al (2018) In situ generation, metabolism and immunomodulatory signaling actions of nitro-conjugated linoleic acid in a murine model of inflammation. Redox Biol 15:522–531. https://doi.org/10.1016/j.redox.2018.01.005
Viniegra A, Goldberg H, Çil Ç, Fine N, Sheikh Z, Galli M, et al (2018) Resolving macrophages counter osteolysis by anabolic actions on bone cells. J Dent Res 97:1160–1169. https://doi.org/10.1177/0022034518777973.
Williams RC (1990) Periodontal disease. N Engl J Med 322:373–381. https://doi.org/10.1056/NEJM199002083220606
Zhou LN, Bi CS, Gao LN, An Y, Chen F, Chen FM (2019) Macrophage polarization in human gingival tissue in response to periodontal disease. Oral Dis 25:265–273. https://doi.org/10.1111/odi.12983
Funding
This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (2021R1A2C1008100).
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by JEL, ARL and E-YC. Conceptualization, formal analysis and project administration were performed by S-JK and In SC. The first draft of the manuscript was written by S-JK and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Conflict of interests
The authors have no relevant financial or non-financial interests to disclose.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Lee, J.E., Lee, A.R., Choi, EY. et al. Effect of nitro-conjugated linoleic acid on the inflammatory response of murine macrophages activated with lipopolysaccharide derived from Prevotella intermedia. Inflammopharmacol 32, 561–573 (2024). https://doi.org/10.1007/s10787-023-01340-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10787-023-01340-8