Protective Effects of Fermented Soymilk Extract on High Glucose-Induced Oxidative Stress in Human Umbilical Vein Endothelial Cells

  • Yi, Na-Ri (Department of Food Science and Nutrition, Pusan National University) ;
  • Park, Min-Jung (Department of Food Science and Nutrition, Pusan National University) ;
  • Han, Ji-Sook (Department of Food Science and Nutrition, Pusan National University)
  • Published : 2010.03.31


We investigated whether the fermented soymilk extract (FSE) has protective effects against high glucose-induced oxidative stress in human umbilical vein endothelial cells (HUVECs). FSE was prepared via fermentation of soymilk with Bacillus subtilis followed by methanol extraction. To determine the protective effect of FSE, oxidative stress was induced by exposing of HUVECs to the high glucose (30 mM) for 48 hr. Exposure of HUVECs to high glucose for 48 hr resulted in a significant (p<0.05) decrease in cell viability, catalase, SOD and GSH-px activity and a significant (p<0.05) increase in intracellular ROS level and thiobarbituric acid reactive substances (TBARS) formation in comparison to the cells treated with 5.5 mM glucose. However, at concentration of 0.1 mg/mL, FSE treatment decreased intracellular ROS level and TBARS formation, and increased cell viability and activities of antioxidant enzymes including catalase, SOD and GSH-px in high glucose pretreated HUVEC. These results suggest that FSE may be able to protect HUVECs from high glucose-induced oxidative stress, partially through the antioxidative defense systems.


  1. Gulter RG, Packer LB, Eartum J, Mori A. 1995. Oxidative stress, antioxidants, aging and disease. In Oxidative stress and aging. Birkhauser Verlag, Basel, Switzerland. p 152-163.
  2. Rosen P, Nawroth PP, King G, Moller W, Tritschler HJ, Packer L. 2001. The role of oxidative stress in the onset and progression of diabetes and its complications: a summary of a congress series sponsored by UNESCO-MCBN, the American Diabetes Association and the German Diabetes Society. Diabetes Metab Res Rev 17: 189-212.
  3. Wiernsperger NF. 2003. Oxidative stress: the special case of diabetes. Biofactors 19: 11-18.
  4. Diabetes Control and Complications Trial Research Group. 1993. The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. N Engl J Med 329: 977-986.
  5. Kinoshita JH. 1990. A thirty year journey in the polyol pathway. Exp Eye Res 50: 567-573.
  6. Kuusisto JL, Mykkanen K, Pyorala, Laakso M. 1994. NIDDM and its metabolic control predict heart disease in elderly subjects. Diabetes 43: 960-967.
  7. Stehouwer CAD, Schaper NC. 1996. The pathogenesis of vascular complications of diabetes mellitus: One voice or many? Eur J Clin Invest 26: 535-543.
  8. Celermajer DS. 1997. Endothelial dysfunction: Does it matter? Is it reversible? J Am Coll Cardiol 30: 325-333.
  9. Wang YC, Yu RC, Chou CC. 2006. Antioxidative activities of soymilk fermented with lactic acid bacteria and bifidobacteria. Food Microbiol 23: 128-135.
  10. Anderson JW, Johnstone BM, Cook N. 1995. Meta-analysis of the effects of soy protein intake on serum lipids. N Engl J Med 333: 276-282.
  11. Potter SM. 1998. Soy protein and cardiovascular disease: the impact of bioactive components in soy. Nutr Rev 56: 231-235.
  12. Hermansen K, Sondergaard M, Hoie L, Carstensen M, Brock B. 2001. Beneficial effects of a soy-based dietary supplement on lipid levels and cardiovascular risk markers in type 2 diabetic subjects. Diabetes Care 24: 228-233.
  13. Anderson JB, Anthony M, Messina M, Garner SC. 1999. Effects of phyto-estrogens on tissues. Nutr Res Rev 12: 75-116.
  14. Bingham SA, Atkinson C, Liggins J, Bluck L, Coward A. 1998. Phyto-estrogens: where are we now? Br J Nutr 79: 393-406.
  15. Setchell KDR, Cassidy A. 1999. Dietary isoflavones: biological effects and relevance to human health. J Nutr 129: 758S-767S.
  16. Rebeca FO, Juana F, Rosario M, Henryk Z, Mariusz KP, Halina K, Concepcion V. 2007. Fermentation as a bio process to obtain functional soybean flours. J Agric Food Chem 55: 8872-8979.
  17. Kuo LC, Cheng WY, Wu RY, Huang CJ, Lee KT. 2006. Hydrolysis of black soybean isoflavone glycosides by Bacillus subtilis natto. Appl Microbiol Biotechnol 73: 314-320.
  18. Fautz R, Husen B, Hechenberger C. 1991. Application of the neutral red assay to monolayer cultures of primary hepatocytes: rapid colorimetric viability determination for the unscheduled DNA synthesis test (UDS). Mutat Res 253: 173-179.
  19. Leloup C, Magnan C, Benani A, Bonnet E, Alquier T, Offer G, Carriere A, Periquet A, Fernandez Y, Ktorza A, Casteilla L, Penicaud L. 2006. Mitochondrial reactive oxygen species are required for hypothalamic glucose sensing. Diabetes 55: 2084-2090.
  20. Fraga CG, Leibovita RM, Roeder RG. 1988. Lipid peroxidation measured as thiobarbituric-reactive substances in tissue slices: characterization and comparision with homogenates and microsomes. Free Radic Biol Med 4: 155-161.
  21. D'Agostino P, Ferlazzo V, Milano S, La Rosa M, Di Bella G, Caruso R. 2001. Anti-inflammatory effects of chemically modified tetracyclines by the inhibition of nitric oxide and interleukin-12 synthesis in J774 cell line. Int Immunopharmacol 1: 1765-1776.
  22. Bradford MM. 1976. A rapid and sensitive method for the quantification of microgram quantities of proteins utilizing the principle of protein-dye binding. Ann Biochem 72: 248-254.
  23. Marklund S, Marklund G. 1974. Involvement of the superoxide anion radical in antioxidant of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47: 469-474.
  24. Aebi H. 1984. Catalase in vitro. Methods Enzymol 105: 121-126.
  25. Lawrence RA, Burk RF. 1976. Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 71: 952-958.
  26. Finkel T, Holbrook NJ. 2000. Oxidants, oxidative stress and the biology of aging. Nature 408: 239-247.
  27. Zheng Z, Chen H, Ke G, Fan Y, Zou H, Sun X, Gu Q, Xu X, Ho PC. 2009. Protective effect of perindopril on diabetic retinopathy is associated with decreased vascular endothelial growth factor-to pigment epithelium-derived factor ratio. Diabetes 58: 954-964.
  28. Sevanian A, Hochstein P. 1985. Mechanism and consequence of lipid peroxidation in biological systems. Annu Rev Nutr 5: 365-390
  29. Hochstein P, Jain SK. 1981. Association of lipid peroxidation and polymerization of membrane proteins with erythrocyte aging. Fed Proc 40: 183-188.
  30. Pacifici RE, Davies KJ. 1991. Protein, lipid and DNA repair systems in oxidative stress: The free radical theory of aging revisited. Gerontology 37: 166-180.
  31. Yen GC, Lai HH. 2002. Inhibitory effects of isoflavones on nitric oxide- or peroxynitrite-mediated DNA damage in RAW 264.7 cells. Food Chem Toxicol 40: 1433-1440.
  32. Mizutani K, Ikeda K, Nishikata T, Yamori Y. 2000. Phytoestrogens attenuate oxidative DNA damage in vascular smooth muscle cells from stroke-prone spontaneously hypertensive rats. J Hypertens 18: 1833-1840.
  33. Mandrup-Poulsen T, Helqvist S, Wogensen LD, Molvig J, Pociot F, Johannesen J, Nerup J. 1990. Cytokine and free radicals as effector molecules in the destruction of pancreatic beta cell. Curr Top Microbiol Immunol 164: 169-193.
  34. Husain K, Somani SM. 1998. Interation of exercise training and chronic ethanol ingestion on testicular antioxidant system in rat. J Appl Toxicol 18: 421-429.<421::AID-JAT532>3.0.CO;2-R
  35. Alptekin N, Seckin S, Yelkenci T, Toker NK, Toker G, Uysal M. 1996. Lipid peroxides, glutathione, $\gamma$-glutamylcysteine synthetase and $\gamma$-gutamyltranspeptidase activities in several tissues of rats following water-immersion stress. Pharmacol Res 34: 167-169.

Cited by

  1. Protective Effects of Sasa Borealis Leaves Extract on High Glucose-Induced Oxidative Stress in Human Umbilical Vein Endothelial Cells vol.39, pp.12, 2010,
  2. Protective effect of Cordyceps militaris against high glucose-induced oxidative stress in human umbilical vein endothelial cells vol.129, pp.3, 2011,