Effect of saccharin on inflammation in 3T3-L1 adipocytes and the related mechanism

  • Kim, Hye Lin (Department of Food Science and Nutrition, Dankook University) ;
  • Ha, Ae Wha (Department of Food Science and Nutrition, Natural Nutraceuticals Industrialization Research Center, DanKook University) ;
  • Kim, Woo Kyoung (Department of Food Science and Nutrition, Dankook University)
  • Received : 2019.03.04
  • Accepted : 2019.12.11
  • Published : 2020.04.01


BACKGROUND/OBJECTIVES: Excessive intake of simple sugars induces obesity and increases the risk of inflammation. Thus, interest in alternative sweeteners as a sugar substitute is increasing. The purpose of this study was to determine the effect of saccharin on inflammation in 3T3-L1 adipocytes. MATERIALS/METHODS: 3T3-L1 preadipocytes were differentiated into adipocytes. The adipocytes were treated with saccharin (0, 50, 100, and 200 ㎍/mL) for 24 h. Inflammation was induced by exposure of treated adipocytes to lipopolysaccharide (LPS) for 18 h and cell proliferation was measured. The concentration of nitric oxide (NO) was measured by using Griess reagent. Protein expressions of nuclear factor kappa B (NF-κB) and inhibitor κB (IκB) were determined by western blot analysis. The mRNA expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin 1β (IL-1β), interleukin 6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor-α (TNF-α) were determined by real-time PCR. RESULTS: Compared with the control group, the amount of NO and the mRNA expression of iNOS in the LPS-treated group were increased by about 17.6% and 46.9%, respectively, (P < 0.05), and those parameter levels were significantly decreased by saccharin treatment (P < 0.05). Protein expression of NF-κB was decreased and that of IκB was increased by saccharin treatment (P < 0.05). Saccharin decreased the mRNA expression of COX-2 and the inflammation cytokines (IL-1β, IL-6, MCP-1, and TNF-α) (P < 0.05). CONCLUSIONS: The results of this study suggest that saccharin can inhibit LPS-induced inflammatory responses in 3T3-L1 adipocytes via the NF-κB pathway.


Supported by : Dankook University


  1. Aller EE, Abete I, Astrup A, Martinez JA, van Baak MA. Starches, sugars and obesity. Nutrients 2011;3:341-69.
  2. Ludwig DS, Peterson KE, Gortmaker SL. Relation between consumption of sugar-sweetened drinks and childhood obesity: a prospective, observational analysis. Lancet 2001;357:505-8.
  3. Hu FB, Malik VS. Sugar-sweetened beverages and risk of obesity and type 2 diabetes: epidemiologic evidence. Physiol Behav 2010;100:47-54.
  4. Beilharz JE, Maniam J, Morris MJ. Short-term exposure to a diet high in fat and sugar, or liquid sugar, selectively impairs hippocampaldependent memory, with differential impacts on inflammation. Behav Brain Res 2016;306:1-7.
  5. Schaffler A, Muller-Ladner U, Scholmerich J, Buchler C. Role of adipose tissue as an inflammatory organ in human diseases. Endocr Rev 2006;27:449-67.
  6. Fantuzzi G. Adipose tissue, adipokines, and inflammation. J Allergy Clin Immunol 2005;115:911-9.
  7. Oeckinghaus A, Ghosh S. The NF-kappaB family of transcription factors and its regulation. Cold Spring Harb Perspect Biol 2009;1:a000034.
  8. Chattopadhyay S, Raychaudhuri U, Chakraborty R. Artificial sweeteners - a review. J Food Sci Technol 2014;51:611-21.
  9. Ju DL. The efficacy and safety of non-nutritive sweeteners. J Korean Diabetes 2015;16:281-6.
  10. Mondal S, Brankow DW, Heidelberger C. Enhancement of oncogenesis in C3H/10T1/2 mouse embryo cell cultures by saccharin. Science 1978;201:1141-2.
  11. Chowaniec J, Hicks RM. Response of the rat to saccharin with particular reference to the urinary bladder. Br J Cancer 1979;39:355-75.
  12. Mortensen A. Sweeteners permitted in the European Union: safety aspects. Scand J Food Nutr 2006;50:104-16.
  13. Kim JW, Baek HH. Safety of saccharin and its current status of regulation in the World. Korean J Food Sci Technol 2011;43:659-74.
  14. Cohen SM, Arnold LL, Emerson JL. Safety of saccharin. Agro Food Ind Hi Tech 2008;19:24-8.
  15. Choi JS, Park SY, Yang MG, Lee DB, Lee TB, Heo JH, Lee MW, Kim SW. Estimation of anti-proliferative activity of saccharin against various cancer cell lines and MSCs. Korean J Clin Lab Sci 2016;48:169-75.
  16. Park JM, Song MK, Kim YJ, Kim YJ. Effect of saccharin intake in restraint-induced stress response reduction in rats. J Korean Biol Nurs Sci 2016;18:36-42.
  17. Lee LS. Saccharin and cyclamate inhibit binding of epidermal growth factor. Proc Natl Acad Sci U S A 1981;78:1042-6.
  18. Thompson MM, Mayer J. Hypoglycemic effects of saccharin in experimental animals. Am J Clin Nutr 1959;7:80-5.
  19. Bailey CJ, Day C, Knapper JM, Turner SL, Flatt PR. Antihyperglycaemic effect of saccharin in diabetic ob/ob mice. Br J Pharmacol 1997;120:74-8.
  20. Csakai A, Smith C, Davis E, Martinko A, Coulup S, Yin H. Saccharin derivatives as inhibitors of interferon-mediated inflammation. J Med Chem 2014;57:5348-55.
  21. Glushakova O, Kosugi T, Roncal C, Mu W, Heinig M, Cirillo P, Sanchez-Lozada LG, Johnson RJ, Nakagawa T. Fructose induces the inflammatory molecule ICAM-1 in endothelial cells. J Am Soc Nephrol 2008;19:1712-20.
  22. DiNicolantonio JJ, Mehta V, Onkaramurthy N, O'Keefe JH. Fructoseinduced inflammation and increased cortisol: a new mechanism for how sugar induces visceral adiposity. Prog Cardiovasc Dis 2018;61:3-9.
  23. Cullberg KB, Larsen JO, Pedersen SB, Richelsen B. Effects of LPS and dietary free fatty acids on MCP-1 in 3T3-L1 adipocytes and macrophages in vitro. Nutr Diabetes 2014;4:e113.
  24. Dobashi K, Asayama K, Nakane T, Kodera K, Hayashibe H, Nakazawa S. Troglitazone inhibits the expression of inducible nitric oxide synthase in adipocytes in vitro and in vivo study in 3T3-L1 cells and Otsuka Long-Evans Tokushima Fatty rats. Life Sci 2000;67:2093-101.
  25. Brasier AR. The nuclear factor-kappaB-interleukin-6 signalling pathway mediating vascular inflammation. Cardiovasc Res 2010;86:211-8.
  26. Nakajima S, Kitamura M. Bidirectional regulation of $NF-{\kappa}B$ by reactive oxygen species: a role of unfolded protein response. Free Radic Biol Med 2013;65:162-74.
  27. Boonkaewwan C, Burodom A. Anti-inflammatory and immunomodulatory activities of stevioside and steviol on colonic epithelial cells. J Sci Food Agric 2013;93:3820-5.
  28. Legler DF, Bruckner M, Uetz-von Allmen E, Krause P. Prostaglandin E2 at new glance: novel insights in functional diversity offer therapeutic chances. Int J Biochem Cell Biol 2010;42:198-201.
  29. Lee W, Kang N, Park SY, Cheong SH, Chang KJ, Kim SH, Um JH, Han EJ, Kim EA, Jeon YJ, Ahn G. Xylose-taurine reduced suppresses the inflammatory responses in lipopolysaccharide-stimulated Raw264.7 macrophages. Adv Exp Med Biol 2017;975:633-42.
  30. Ballak DB, Stienstra R, Tack CJ, Dinarello CA, van Diepen JA. IL-1 family members in the pathogenesis and treatment of metabolic disease: Focus on adipose tissue inflammation and insulin resistance. Cytokine 2015;75:280-90.
  31. Su C, Chen M, Huang H, Lin J. Testosterone enhances lipopolysaccharide-induced interleukin-6 and macrophage chemotactic protein-1 expression by activating the extracellular signal-regulated kinase 1/2/nuclear $factor-{\kappa}B$ signalling pathways in 3T3-L1 adipocytes. Mol Med Rep 2015;12:696-704.
  32. Blancas-Flores G, Alarcon-Aguilar FJ, Garcia-Macedo R, Almanza-Perez JC, Flores-Saenz JL, Roman-Ramos R, Ventura-Gallegos JL, Kumate J, Zentella-Dehesa A, Cruz M. Glycine suppresses $TNF-{\alpha}$-induced activation of $NF-{\kappa}B$ in differentiated 3T3-L1 adipocytes. Eur J Pharmacol 2012;689:270-7.
  33. Kim SY, Jo MJ, Hwangbo M, Back YD, Jeong TY, Cho IJ, Jee SY. Anti-inflammatory effect of Stevia rebaudiana as a result of $NF-{\kappa}B$ and MAPK inhibition. J Korean Med Ophthalmol Otolaryngol Dermatol 2013;26:54-64.
  34. Wang Z, Xue L, Guo C, Han B, Pan C, Zhao S, Song H, Ma Q. Stevioside ameliorates high-fat diet-induced insulin resistance and adipose tissue inflammation by downregulating the $NF-{\kappa}B$ pathway. Biochem Biophys Res Commun 2012;417:1280-5.