The micosporine-like amino acids-rich aqueous methanol extract of laver (Porphyra yezoensis) inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes

  • Kim, Hyunhee (Department of Chemistry, Incheon National University) ;
  • Lee, Yunjung (Department of Chemistry, Incheon National University) ;
  • Han, Taejun (Department of Marine Sciences, Incheon National University) ;
  • Choi, Eun-Mi (Department of Chemistry, Incheon National University)
  • Received : 2015.03.31
  • Accepted : 2015.06.16
  • Published : 2015.12.01


BACKGROUND/OBJECTIVES: Increased mass of adipose tissue in obese persons is caused by excessive adipogenesis, which is elaborately controlled by an array of transcription factors. Inhibition of adipogenesis by diverse plant-derived substances has been explored. The aim of the current study was to examine the effects of the aqueous methanol extract of laver (Porphyra yezoensis) on adipogenesis and apoptosis in 3T3-L1 adipocytes and to investigate the mechanism underlying the effect of the laver extract. MATERIALS/METHODS: 3T3-L1 cells were treated with various concentrations of laver extract in differentiation medium. Lipid accumulation, expression of adipogenic proteins, including CCAAT enhancer-binding protein ${\alpha}$, peroxisome proliferator-activated receptor ${\gamma}$, fatty acid binding protein 4, and fatty acid synthase, cell viability, apoptosis, and the total content and the ratio of reduced to oxidized forms of glutathione (GSH/GSSG) were analyzed. RESULTS: Treatment with laver extract resulted in a significant decrease in lipid accumulation in 3T3-L1 adipocytes, which showed correlation with a reduction in expression of adipogenic proteins. Treatment with laver extract also resulted in a decrease in the viability of preadipocytes and an increase in the apoptosis of mature adipocytes. Treatment with laver extract led to exacerbated depletion of cellular glutathione and abolished the transient increase in GSH/GSSG ratio during adipogenesis in 3T3-L1 adipocytes. CONCLUSION: Results of our study demonstrated that treatment with the laver extract caused inhibition of adipogenesis, a decrease in proliferation of preadipocytes, and an increase in the apoptosis of mature adipocytes. It appears that these effects were caused by increasing oxidative stress, as demonstrated by the depletion and oxidation of the cellular glutathione pool in the extract-treated adipocytes. Our results suggest that a prooxidant role of laver extract is associated with its antiadipogenic and proapoptotic effects.


Laver extract;adipogenesis;apoptosis;oxidative stress;glutathione


Grant : 뷰티산업 글로컬 리더 육성 사업단


  1. Saltiel AR, Kahn CR. Insulin signalling and the regulation of glucose and lipid metabolism. Nature 2001;414:799-806.
  2. Camp HS, Ren D, Leff T. Adipogenesis and fat-cell function in obesity and diabetes. Trends Mol Med 2002;8:442-7.
  3. Couillard C, Mauriege P, Imbeault P, Prud'homme D, Nadeau A, Tremblay A, Bouchard C, Despres JP. Hyperleptinemia is more closely associated with adipose cell hypertrophy than with adipose tissue hyperplasia. Int J Obes Relat Metab Disord 2000;24:782-8.
  4. Farmer SR. Transcriptional control of adipocyte formation. Cell Metab 2006;4:263-73.
  5. Lowe CE, O'Rahilly S, Rochford JJ. Adipogenesis at a glance. J Cell Sci 2011;124:2681-6.
  6. Gregoire FM, Smas CM, Sul HS. Understanding adipocyte differentiation. Physiol Rev 1998;78:783-809.
  7. Rayalam S, Della-Fera MA, Baile CA. Phytochemicals and regulation of the adipocyte life cycle. J Nutr Biochem 2008;19:717-26.
  8. Lee JC, Hou MF, Huang HW, Chang FR, Yeh CC, Tang JY, Chang HW. Marine algal natural products with anti-oxidative, anti-inflammatory, and anti-cancer properties. Cancer Cell Int 2013;13:55.
  9. Faulkner DJ. Marine natural products. Nat Prod Rep 2002;19:1-48.
  10. Yuan YV, Walsh NA. Antioxidant and antiproliferative activities of extracts from a variety of edible seaweeds. Food Chem Toxicol 2006;44:1144-50.
  11. Groniger A, Sinha RP, Klisch M, Hader DP. Photoprotective compounds in cyanobacteria, phytoplankton and macroalgae--a database. J Photochem Photobiol B 2000;58:115-22.
  12. Sinha RP, Singh SP, Hader DP. Database on mycosporines and mycosporine-like amino acids (MAAs) in fungi, cyanobacteria, macroalgae, phytoplankton and animals. J Photochem Photobiol B 2007;89:29-35.
  13. Esteban R, Martinez B, Fernandez-Marin B, Maria Becerril J, Garcia-Plazaola JI. Carotenoid composition in Rhodophyta: insights into xanthophyll regulation in Corallina elongata. Eur J Phycol 2009;44:221-30.
  14. Olsen EK, Hansen E, Isaksson J, Andersen JH. Cellular antioxidant effect of four bromophenols from the red algae, Vertebrata lanosa. Mar Drugs 2013;11:2769-84.
  15. Wang H, Fu XM, Han CC. The potential applications of marine bioactives against diabetes and obesity. Am J Mar Sci 2014;2:1-8.
  16. Kim KJ, Lee OH, Lee BY. Fucoidan, a sulfated polysaccharide, inhibits adipogenesis through the mitogen-activated protein kinase pathway in 3T3-L1 preadipocytes. Life Sci 2010;86:791-7.
  17. Maeda H, Hosokawa M, Sashima T, Takahashi N, Kawada T, Miyashita K. Fucoxanthin and its metabolite, fucoxanthinol, suppress adipocyte differentiation in 3T3-L1 cells. Int J Mol Med 2006;18:147-52.
  18. Jeon HJ, Seo MJ, Choi HS, Lee OH, Lee BY. Gelidium elegans, an edible red seaweed, and hesperidin inhibit lipid accumulation and production of reactive oxygen species and reactive nitrogen species in 3T3-L1 and RAW264.7 cells. Phytother Res 2014;28:1701-9.
  19. Kim HM, Kang SI, Shin HS, Ko HC, Hong YS, Kang SW, Yoon SA, Kim JH, Kim SJ. Anti-obesity effect of Komulkosiraegi [Gracilaria vermiculophylla (Ohmi) Papenfuss] extract in 3T3-L1 cells. Food Sci Biotechnol 2012;21:83-9.
  20. Gironi F, Piemonte V. Temperature and solvent effects on polyphenol extraction process from chestnut tree wood. Chem Eng Res Des 2011;89:857-62.
  21. Volkmann M, Gorbushina AA. A broadly applicable method for extraction and characterization of mycosporines and mycosporine-like amino acids of terrestrial, marine and freshwater origin. FEMS Microbiol Lett 2006;255:286-95.
  22. Klisch M, Hader DP. Wavelength dependence of mycosporine-like amino acid synthesis in Gyrodinium dorsum. J Photochem Photobiol B 2002;66:60-6.
  23. Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am K Enol Vitic 1965;16:144-58.
  24. Ramirez-Zacarias JL, Castro-Munozledo F, Kuri-Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil red O. Histochemistry 1992;97:493-7.
  25. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
  26. Neuschwander-Tetri BA, Roll FJ. Glutathione measurement by high-performance liquid chromatography separation and fluorometric detection of the glutathione-orthophthalaldehyde adduct. Anal Biochem 1989;179:236-41.
  27. Kim S, You DH, Han T, Choi EM. Modulation of viability and apoptosis of UVB-exposed human keratinocyte HaCaT cells by aqueous methanol extract of laver (Porphyra yezoensis). J Photochem Photobiol B 2014;141:301-7.
  28. Zubia M, Fabre MS, Kerjean V, Deslandes E. Antioxidant and cytotoxic activities of some red algae (Rhodophyta) from Brittany coasts (France). Bot Mar 2009;52:268-77.
  29. Hwang ES, Thi ND. Effects of extraction and processing methods on antioxidant compound contents and radical scavenging activities of laver (Porphyra tenera). Prev Nutr Food Sci 2014;19:40-8.
  30. Landete JM. Updated knowledge about polyphenols: functions, bioavailability, metabolism, and health. Crit Rev Food Sci Nutr 2012;52:936-48.
  31. Martin-Fernandez C, Bales J, Hodgkinson C, Welman A, Welham MJ, Dive C, Morrow CJ. Blocking phosphoinositide 3-kinase activity in colorectal cancer cells reduces proliferation but does not increase apoptosis alone or in combination with cytotoxic drugs. Mol Cancer Res 2009;7:955-65.
  32. Hamtiaux L, Hansoulle L, Dauguet N, Muccioli GG, Gallez B, Lambert DM. Increasing antiproliferative properties of endocannabinoids in N1E-115 neuroblastoma cells through inhibition of their metabolism. PLoS One 2011;6:e26823.
  33. Rayalam S, Yang JY, Ambati S, Della-Fera MA, Baile CA. Resveratrol induces apoptosis and inhibits adipogenesis in 3T3-L1 adipocytes. Phytother Res 2008;22:1367-71.
  34. Yang JY, Della-Fera MA, Rayalam S, Ambati S, Hartzell DL, Park HJ, Baile CA. Enhanced inhibition of adipogenesis and induction of apoptosis in 3T3-L1 adipocytes with combinations of resveratrol and quercetin. Life Sci 2008;82:1032-9.
  35. Kim H, Hiraishi A, Tsuchiya K, Sakamoto K. (-) Epigallocatechin gallate suppresses the differentiation of 3T3-L1 preadipocytes through transcription factors FoxO1 and SREBP1c. Cytotechnology 2010;62:245-55.
  36. Andersen C, Rayalam S, Della-Fera MA, Baile CA. Phytochemicals and adipogenesis. Biofactors 2010;36:415-22.
  37. Liu GS, Chan EC, Higuchi M, Dusting GJ, Jiang F. Redox mechanisms in regulation of adipocyte differentiation: beyond a general stress response. Cells 2012;1:976-93.
  38. Gummersbach C, Hemmrich K, Kröncke KD, Suschek CV, Fehsel K, Pallua N. New aspects of adipogenesis: radicals and oxidative stress. Differentiation 2009;77:115-20.
  39. Mytilineou C, Kramer BC, Yabut JA. Glutathione depletion and oxidative stress. Parkinsonism Relat Disord 2002;8:385-7.
  40. Torres-Ramirez N, Baiza-Gutman LA, Garcia-Macedo R, Ortega-Camarillo C, Contreras-Ramos A, Medina-Navarro R, Cruz M, Ibanez-Hernandez MA, Diaz-Flores M. Nicotinamide, a glucose-6-phosphate dehydrogenase non-competitive mixed inhibitor, modifies redox balance and lipid accumulation in 3T3-L1 cells. Life Sci 2013;93:975-85.
  41. Ducluzeau PH, Priou M, Weitheimer M, Flamment M, Duluc L, Iacobazi F, Soleti R, Simard G, Durand A, Rieusset J, Andriantsitohaina R, Malthiery Y. Dynamic regulation of mitochondrial network and oxidative functions during 3T3-L1 fat cell differentiation. J Physiol Biochem 2011;67:285-96.
  42. Maher J, Yamamoto M. The rise of antioxidant signaling--the evolution and hormetic actions of Nrf2. Toxicol Appl Pharmacol 2010;244:4-15.
  43. Hou Y, Xue P, Bai Y, Liu D, Woods CG, Yarborough K, Fu J, Zhang Q, Sun G, Collins S, Chan JY, Yamamoto M, Andersen ME, Pi J. Nuclear factor erythroid-derived factor 2-related factor 2 regulates transcription of CCAAT/enhancer-binding protein beta during adipogenesis. Free Radic Biol Med 2012;52:462-72.
  44. Findeisen HM, Pearson KJ, Gizard F, Zhao Y, Qing H, Jones KL, Cohn D, Heywood EB, de Cabo R, Bruemmer D. Oxidative stress accumulates in adipose tissue during aging and inhibits adipogenesis. PLoS One 2011;6:e18532.
  45. Tao L, Park JY, Lambert JD. Differential prooxidative effects of the green tea polyphenol, (-)-epigallocatechin-3-gallate, in normal and oral cancer cells are related to differences in sirtuin 3 signaling. Mol Nutr Food Res 2015;59:203-11.
  46. Kucinska M, Piotrowska H, Luczak MW, Mikula-Pietrasik J, Ksiazek K, Wozniak M, Wierzchowski M, Dudka J, Jager W, Murias M. Effects of hydroxylated resveratrol analogs on oxidative stress and cancer cells death in human acute T cell leukemia cell line: prooxidative potential of hydroxylated resveratrol analogs. Chem Biol Interact 2014;209:96-110.
  47. Wang CT, Chang HH, Hsiao CH, Lee MJ, Ku HC, Hu YJ, Kao YH. The effects of green tea (-)-epigallocatechin-3-gallate on reactive oxygen species in 3T3-L1 preadipocytes and adipocytes depend on the glutathione and 67 kDa laminin receptor pathways. Mol Nutr Food Res 2009;53:349-60.