Skip to main content
Top
Published in: Inflammation 4/2013

01-08-2013

Nitric Oxide and Interleukins are Involved in Cell Proliferation of RAW264.7 Macrophages Activated by Viili Exopolysaccharides

Authors: Junhua Wu, Mengxian Li, Ling Liu, Qi An, Jinlu Zhang, Jingkai Zhang, Meiling Li, Weigang Duan, Dequan Liu, Zhenjing Li, Cheng Luo

Published in: Inflammation | Issue 4/2013

Login to get access

Abstract

Viili has been traditionally regarded as healthy food; viili exopolysaccharides (VEPS) function as antioxidants, but the molecular and cellular mechanisms, especially its immune functions, remain largely unclear. To assess VEPS’s immunological roles, VEPS were separated by Sevage’s method and purified by anion exchange chromatography. Cell proliferation, phagocytosis, releases of nitric oxide (NO), interleukin (IL)-1β, and IL-6, the inducible nitric oxide synthase (iNOS) gene expression by reverse transcription polymerase chain reaction (RT-PCR) and iNOS protein by Western blotting, and morphology by scanning electron microscopy in lipopolysaccharides (LPS)/VEPS-stimulated and non-stimulated RAW264.7 macrophages were analyzed. VEPS increased cell proliferation at 50–200 μg/mL. The uptake of neutral red for the indication of phagocytosis and releases of NO, IL-6, and IL-1β were enhanced after exposure to LPS and VEPS. Gene expressions of iNOS, IL-6, and IL-1β and protein expressions of iNOS were increased with VEPS. The RAW264.7 cell treated with VEPS became flattened, a strong indication of the activation of macrophages. We concluded that VEPS promoted the activation of macrophages in which NO, IL-6, and IL-1β were involved; the release of NO and other cytokines may eventually activate lymphocytes, increasing nonspecific (innate) and specific immunity in humans.
Literature
1.
go back to reference Nakajima, H., T. Hirota, T. Toba, T. Itoh, and S. Adachi. 1992. Structure of the extracellular polysaccharide from slime-forming Lactococcus lactis subsp. cremoris SBT 0495. Carbohydrate Research 224: 245–253.PubMedCrossRef Nakajima, H., T. Hirota, T. Toba, T. Itoh, and S. Adachi. 1992. Structure of the extracellular polysaccharide from slime-forming Lactococcus lactis subsp. cremoris SBT 0495. Carbohydrate Research 224: 245–253.PubMedCrossRef
2.
go back to reference Higashimura, M., B.W. Mulder-Bosman, R. Reich, T. Iwasaki, and G.W. Robijn. 2000. Solution properties of viilian, the exopolysaccharide from Lactococcus lactis subsp. cremoris SBT 0495. Biopolymers 54: 143–158.PubMedCrossRef Higashimura, M., B.W. Mulder-Bosman, R. Reich, T. Iwasaki, and G.W. Robijn. 2000. Solution properties of viilian, the exopolysaccharide from Lactococcus lactis subsp. cremoris SBT 0495. Biopolymers 54: 143–158.PubMedCrossRef
3.
go back to reference Kitazawa, H., T. Yamaguchi, M. Miura, T. Saito, and T. Itoh. 1993. B cell mitogen produced by slime-forming, encapsulated Lactococcus lactis ssp. cremoris isolated from ropy sour milk, viili. Journal of Dairy Science 76: 1514–1519.PubMedCrossRef Kitazawa, H., T. Yamaguchi, M. Miura, T. Saito, and T. Itoh. 1993. B cell mitogen produced by slime-forming, encapsulated Lactococcus lactis ssp. cremoris isolated from ropy sour milk, viili. Journal of Dairy Science 76: 1514–1519.PubMedCrossRef
4.
go back to reference Kitazawa, H., M. Tohno, T. Shimosato, and T. Saito. 2007. Development of molecular immunoassay system for probiotics via toll-like receptors based on food immunology. Animal Science Journal 79: 11–21.CrossRef Kitazawa, H., M. Tohno, T. Shimosato, and T. Saito. 2007. Development of molecular immunoassay system for probiotics via toll-like receptors based on food immunology. Animal Science Journal 79: 11–21.CrossRef
5.
go back to reference Kekkonen, R.A., E. Kajasto, M. Miettinen, V. Veckman, R. Korpela, and I. Julkunen. 2008. Probiotic Leuconostoc mesenteroides ssp. cremoris and Streptococcus thermophilus induce IL-12 and IFN-gamma production. World Journal of Gastroenterology 14: 1192–1203.PubMedCrossRef Kekkonen, R.A., E. Kajasto, M. Miettinen, V. Veckman, R. Korpela, and I. Julkunen. 2008. Probiotic Leuconostoc mesenteroides ssp. cremoris and Streptococcus thermophilus induce IL-12 and IFN-gamma production. World Journal of Gastroenterology 14: 1192–1203.PubMedCrossRef
6.
go back to reference Liu, L., J.H. Wu, J.L. Zhang, Z.J. Li, C.L. Wang, M.H. Chen, Y.R. Wang, Y.X. Sun, L.K. Wang, and C. Luo. 2011. A compatibility assay of ursolic acid and foodborne microbial exopolysaccharides by antioxidant power and anti-proliferative properties in hepatocarcinoma cells. Journal of Food, Agriculture & Environment 10: 111–1114. Liu, L., J.H. Wu, J.L. Zhang, Z.J. Li, C.L. Wang, M.H. Chen, Y.R. Wang, Y.X. Sun, L.K. Wang, and C. Luo. 2011. A compatibility assay of ursolic acid and foodborne microbial exopolysaccharides by antioxidant power and anti-proliferative properties in hepatocarcinoma cells. Journal of Food, Agriculture & Environment 10: 111–1114.
7.
go back to reference Lee, M.Y., J.A. Lee, C.S. Seo, H. Ha, H. Lee, J.K. Son, and H.K. Shin. 2011. Anti-inflammatory activity of Angelica dahurica ethanolic extract on RAW264.7 cells via upregulation of heme oxygenase-1. Food and Chemical Toxicology 49: 1047–1055.PubMedCrossRef Lee, M.Y., J.A. Lee, C.S. Seo, H. Ha, H. Lee, J.K. Son, and H.K. Shin. 2011. Anti-inflammatory activity of Angelica dahurica ethanolic extract on RAW264.7 cells via upregulation of heme oxygenase-1. Food and Chemical Toxicology 49: 1047–1055.PubMedCrossRef
8.
go back to reference Shin, H.Y., C.H. Shin, T.Y. Shin, E.J. Lee, and H.M. Kim. 2003. Effect of bojungikki-tang on lipopolysaccharide-induced cytokine production from peripheral blood mononuclear cells of chronic fatigue syndrome patients. Immunopharmacology and Immunotoxicology 25: 491–501.PubMedCrossRef Shin, H.Y., C.H. Shin, T.Y. Shin, E.J. Lee, and H.M. Kim. 2003. Effect of bojungikki-tang on lipopolysaccharide-induced cytokine production from peripheral blood mononuclear cells of chronic fatigue syndrome patients. Immunopharmacology and Immunotoxicology 25: 491–501.PubMedCrossRef
9.
go back to reference Kröncke, K.D., K. Fehsel, and V. Kolb-Bachofen. 1998. Inducible nitric oxide synthase in human diseases. Clinical Experimental Immunology 113: 147–156.CrossRef Kröncke, K.D., K. Fehsel, and V. Kolb-Bachofen. 1998. Inducible nitric oxide synthase in human diseases. Clinical Experimental Immunology 113: 147–156.CrossRef
10.
go back to reference Ohshima, H., and H. Bartsch. 1994. Chronic infections and inflammatory processes as cancer risk factors: Possible role of nitric oxide in carcinogenesis. Mutation Research 305: 253–264.PubMedCrossRef Ohshima, H., and H. Bartsch. 1994. Chronic infections and inflammatory processes as cancer risk factors: Possible role of nitric oxide in carcinogenesis. Mutation Research 305: 253–264.PubMedCrossRef
11.
go back to reference Korhonen, R., A. Lahti, H. Kankaanranta, and E. Moilanen. 2005. Nitric oxide production and signaling in inflammation. Current Drug Targets Inflammation Allergy 4: 471–479.PubMedCrossRef Korhonen, R., A. Lahti, H. Kankaanranta, and E. Moilanen. 2005. Nitric oxide production and signaling in inflammation. Current Drug Targets Inflammation Allergy 4: 471–479.PubMedCrossRef
12.
go back to reference Valko, M., D. Leibfritz, J. Moncol, M.T. Cronin, M. Mazur, and J. Telser. 2007. Free radicals and antioxidants in normal physiological functions and human disease. The International Journal of Biochemistry & Cell Biology 39: 44–84.CrossRef Valko, M., D. Leibfritz, J. Moncol, M.T. Cronin, M. Mazur, and J. Telser. 2007. Free radicals and antioxidants in normal physiological functions and human disease. The International Journal of Biochemistry & Cell Biology 39: 44–84.CrossRef
13.
go back to reference Yang, Z., E. Huttunen, M. Staaf, G. Widmalm, and H. Tenhu. 1999. Separation, purification and characterization of extracellular polysaccharides produced by slime-forming¸ Lactococcus lactis ssp. cremoris strains. International Dairy Journal 9: 631–638.CrossRef Yang, Z., E. Huttunen, M. Staaf, G. Widmalm, and H. Tenhu. 1999. Separation, purification and characterization of extracellular polysaccharides produced by slime-forming¸ Lactococcus lactis ssp. cremoris strains. International Dairy Journal 9: 631–638.CrossRef
14.
go back to reference Papis, E., S.J. Davies, and A.N. Jha. 2011. Relative sensitivity of fish and mammalian cells to the antibiotic, trimethoprim: Cytotoxic and genotoxic responses as determined by neutral red retention, Comet and micronucleus assays. Ecotoxicology 20: 208–217.PubMedCrossRef Papis, E., S.J. Davies, and A.N. Jha. 2011. Relative sensitivity of fish and mammalian cells to the antibiotic, trimethoprim: Cytotoxic and genotoxic responses as determined by neutral red retention, Comet and micronucleus assays. Ecotoxicology 20: 208–217.PubMedCrossRef
15.
go back to reference Babich, H., M.R. Palace, and A. Stern. 1993. Oxidative stress in fish cells: In vitro studies. Archives Environmental Contamination Toxicology 24: 173–178.CrossRef Babich, H., M.R. Palace, and A. Stern. 1993. Oxidative stress in fish cells: In vitro studies. Archives Environmental Contamination Toxicology 24: 173–178.CrossRef
16.
go back to reference Markewitz, B.A., J.R. Michael, and D.E. Kohan. 1993. Cytokine-induced expression of a nitric oxide synthase in rat renal tubule cells. Journal of Clinical Investigation 91: 2138–2143.PubMedCrossRef Markewitz, B.A., J.R. Michael, and D.E. Kohan. 1993. Cytokine-induced expression of a nitric oxide synthase in rat renal tubule cells. Journal of Clinical Investigation 91: 2138–2143.PubMedCrossRef
17.
go back to reference Maa, P., H.T. Liua, W. Peng, Q.S. Xua, X.F. Bai, Y.G. Du, and C. Yu. 2011. Chitosan oligosaccharides inhibit LPS-induced over-expression of IL-6 and TNF-α in RAW264.7 macrophage cells through blockade of mitogen-activated protein kinase (MAPK) and PI3K/Akt signaling pathways. Carbohydrate Polymers 84: 1391–1398.CrossRef Maa, P., H.T. Liua, W. Peng, Q.S. Xua, X.F. Bai, Y.G. Du, and C. Yu. 2011. Chitosan oligosaccharides inhibit LPS-induced over-expression of IL-6 and TNF-α in RAW264.7 macrophage cells through blockade of mitogen-activated protein kinase (MAPK) and PI3K/Akt signaling pathways. Carbohydrate Polymers 84: 1391–1398.CrossRef
18.
go back to reference Liang, Q., Q. Wu, J. Jiang, J. Duan, C. Wang, M.D. Smith, H. Lu, Q. Wang, P. Nagarkatti, and D. Fan. 2011. Characterization of sparstolonin B, a Chinese herb-derived compound, as a selective Toll-like receptor antagonist with potent anti-inflammatory properties. Journal of Biological Chemistry 286: 26470–26479.PubMedCrossRef Liang, Q., Q. Wu, J. Jiang, J. Duan, C. Wang, M.D. Smith, H. Lu, Q. Wang, P. Nagarkatti, and D. Fan. 2011. Characterization of sparstolonin B, a Chinese herb-derived compound, as a selective Toll-like receptor antagonist with potent anti-inflammatory properties. Journal of Biological Chemistry 286: 26470–26479.PubMedCrossRef
19.
go back to reference Kousteni, S., T. Bellido, L.I. Plotkin, C.A. O’Brien, D.L. Bodenner, L. Han, K. Han, G.B. DiGregorio, J.A. Katzenellenbogen, B.S. Katzenellenbogen, P.K. Roberson, R.S. Weinstein, R.L. Jilka, and S.C. Manolagas. 2001. Nongenotropic, sex-nonspecific signaling through the estrogen or androgen receptors: Dissociation from transcriptional activity. Cell 104: 719–730.PubMed Kousteni, S., T. Bellido, L.I. Plotkin, C.A. O’Brien, D.L. Bodenner, L. Han, K. Han, G.B. DiGregorio, J.A. Katzenellenbogen, B.S. Katzenellenbogen, P.K. Roberson, R.S. Weinstein, R.L. Jilka, and S.C. Manolagas. 2001. Nongenotropic, sex-nonspecific signaling through the estrogen or androgen receptors: Dissociation from transcriptional activity. Cell 104: 719–730.PubMed
20.
go back to reference Tohno, M., H. Kitazawa, T. Shimosato, M. Matsumoto, S. Katoh, Y. Kawai, and T. Saito. 2005. A swine toll-like receptor 2-expressing transfectant as a potential primary screening system for immunobiotic microorganisms. FEMS Immunology Medical Microbiology 44: 283–288.CrossRef Tohno, M., H. Kitazawa, T. Shimosato, M. Matsumoto, S. Katoh, Y. Kawai, and T. Saito. 2005. A swine toll-like receptor 2-expressing transfectant as a potential primary screening system for immunobiotic microorganisms. FEMS Immunology Medical Microbiology 44: 283–288.CrossRef
21.
go back to reference Tohno, M., T. Shimazu, W. Ueda, D. Anzawa, H. Aso, J. Nishimura, Y. Kawai, Y. Saito, T. Saito, and H. Kitazawa. 2007. Molecular cloning of porcine RP105/MD-1 involved in recognition of extracellular phosphopolysaccharides from Lactococcus lactis ssp. cremoris. Molecular Immunology 44: 2566–2577.PubMedCrossRef Tohno, M., T. Shimazu, W. Ueda, D. Anzawa, H. Aso, J. Nishimura, Y. Kawai, Y. Saito, T. Saito, and H. Kitazawa. 2007. Molecular cloning of porcine RP105/MD-1 involved in recognition of extracellular phosphopolysaccharides from Lactococcus lactis ssp. cremoris. Molecular Immunology 44: 2566–2577.PubMedCrossRef
22.
go back to reference Lyons, C.R., G.J. Orloff, and J.M. Cunningham. 1992. Molecular cloning and functional expression of an inducible nitric oxide synthase from a murine macrophage cell line. The Journal of Biological Chemistry 267: 6370–6374.PubMed Lyons, C.R., G.J. Orloff, and J.M. Cunningham. 1992. Molecular cloning and functional expression of an inducible nitric oxide synthase from a murine macrophage cell line. The Journal of Biological Chemistry 267: 6370–6374.PubMed
23.
go back to reference Jin, M., S.J. Suh, J.H. Yang, Y. Lu, S.J. Kim, S. Kwon, T.H. Jo, J.W. Kim, Y.I. Park, G.W. Ahn, C.K. Lee, C.H. Kim, J.K. Son, K.H. Son, and H.W. Chang. 2010. Anti-inflammatory activity of bark of Dioscorea batatas DECNE through the inhibition of iNOS and COX-2 expressions in RAW264.7 cells via NF-κB and ERK1/2 inactivation. Food and Chemical Toxicology 48: 3073–3079.PubMedCrossRef Jin, M., S.J. Suh, J.H. Yang, Y. Lu, S.J. Kim, S. Kwon, T.H. Jo, J.W. Kim, Y.I. Park, G.W. Ahn, C.K. Lee, C.H. Kim, J.K. Son, K.H. Son, and H.W. Chang. 2010. Anti-inflammatory activity of bark of Dioscorea batatas DECNE through the inhibition of iNOS and COX-2 expressions in RAW264.7 cells via NF-κB and ERK1/2 inactivation. Food and Chemical Toxicology 48: 3073–3079.PubMedCrossRef
24.
go back to reference Manucha, W., L. Oliveros, L. Carrizo, A. Seltzer, and P. Vallés. 2004. Losartan modulation on NOS isoforms and COX-2 expression in early renal fibrogenesis in unilateral obstruction. Kidney International 65: 2091–2107.PubMedCrossRef Manucha, W., L. Oliveros, L. Carrizo, A. Seltzer, and P. Vallés. 2004. Losartan modulation on NOS isoforms and COX-2 expression in early renal fibrogenesis in unilateral obstruction. Kidney International 65: 2091–2107.PubMedCrossRef
25.
go back to reference Luo, C., T. Vooder, E. Urgard, and A. Metspalu. 2011. The role of COX-2 and Nrf2/ARE in anti-inflammation and antioxidative stress: Aging and anti-aging. Med Hypotheses 77: 174–178.PubMedCrossRef Luo, C., T. Vooder, E. Urgard, and A. Metspalu. 2011. The role of COX-2 and Nrf2/ARE in anti-inflammation and antioxidative stress: Aging and anti-aging. Med Hypotheses 77: 174–178.PubMedCrossRef
26.
go back to reference Luo, C., M. Kallajoki, R. Gross, M. Mulari, T. Teros, L. Ylinen, M. Mäkinen, J. Laine, and O. Simell. 2002. Cellular distribution and contribution of cyclooxygenase COX-2 to diabetogenesis in NOD mouse. Cell Tissue Research 310: 169–175.PubMedCrossRef Luo, C., M. Kallajoki, R. Gross, M. Mulari, T. Teros, L. Ylinen, M. Mäkinen, J. Laine, and O. Simell. 2002. Cellular distribution and contribution of cyclooxygenase COX-2 to diabetogenesis in NOD mouse. Cell Tissue Research 310: 169–175.PubMedCrossRef
27.
go back to reference Mogensen, T.H. 2009. Pathogen recognition and inflammatory signaling in innate immune defenses. Clinical Microbiology Reviews 22: 240–273.PubMedCrossRef Mogensen, T.H. 2009. Pathogen recognition and inflammatory signaling in innate immune defenses. Clinical Microbiology Reviews 22: 240–273.PubMedCrossRef
28.
go back to reference Schmidt, A., E. Caron, and A. Hall. 2001. Lipopolysaccharide-induced activation of beta2-integrin function in macrophages requires Irak kinase activity, p38 mitogen-activated protein kinase, and the Rap1 GTPase. Molecular and Cellular Biology 21: 438–448.PubMedCrossRef Schmidt, A., E. Caron, and A. Hall. 2001. Lipopolysaccharide-induced activation of beta2-integrin function in macrophages requires Irak kinase activity, p38 mitogen-activated protein kinase, and the Rap1 GTPase. Molecular and Cellular Biology 21: 438–448.PubMedCrossRef
29.
go back to reference Begum, R., M.S. Nur-E-Kamal, and M.A. Zaman. 2004. The role of Rho GTPases in the regulation of the rearrangement of actin cytoskeleton and cell movement. Experimental & Molecular Medicine 36: 358–366.CrossRef Begum, R., M.S. Nur-E-Kamal, and M.A. Zaman. 2004. The role of Rho GTPases in the regulation of the rearrangement of actin cytoskeleton and cell movement. Experimental & Molecular Medicine 36: 358–366.CrossRef
Metadata
Title
Nitric Oxide and Interleukins are Involved in Cell Proliferation of RAW264.7 Macrophages Activated by Viili Exopolysaccharides
Authors
Junhua Wu
Mengxian Li
Ling Liu
Qi An
Jinlu Zhang
Jingkai Zhang
Meiling Li
Weigang Duan
Dequan Liu
Zhenjing Li
Cheng Luo
Publication date
01-08-2013
Publisher
Springer US
Published in
Inflammation / Issue 4/2013
Print ISSN: 0360-3997
Electronic ISSN: 1573-2576
DOI
https://doi.org/10.1007/s10753-013-9626-y

Other articles of this Issue 4/2013

Inflammation 4/2013 Go to the issue
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine