Genes & Nutrition

, Volume 7, Issue 3, pp 459–469 | Cite as

Benfotiamine increases glucose oxidation and downregulates NADPH oxidase 4 expression in cultured human myotubes exposed to both normal and high glucose concentrations

  • D. A. Fraser
  • N. P. Hessvik
  • N. Nikolić
  • V. Aas
  • K. F. Hanssen
  • S. K. Bøhn
  • G. H. Thoresen
  • A. C. Rustan
Research Paper


The aim of the present work was to study the effects of benfotiamine (S-benzoylthiamine O-monophosphate) on glucose and lipid metabolism and gene expression in differentiated human skeletal muscle cells (myotubes) incubated for 4 days under normal (5.5 mM glucose) and hyperglycemic (20 mM glucose) conditions. Myotubes established from lean, healthy volunteers were treated with benfotiamine for 4 days. Glucose and lipid metabolism were studied with labeled precursors. Gene expression was measured using real-time polymerase chain reaction (qPCR) and microarray technology. Benfotiamine significantly increased glucose oxidation under normoglycemic (35 and 49% increase at 100 and 200 μM benfotiamine, respectively) as well as hyperglycemic conditions (70% increase at 200 μM benfotiamine). Benfotiamine also increased glucose uptake. In comparison, thiamine (200 μM) increased overall glucose metabolism but did not change glucose oxidation. In contrast to glucose, mitochondrial lipid oxidation and overall lipid metabolism were unchanged by benfotiamine. The expression of NADPH oxidase 4 (NOX4) was significantly downregulated by benfotiamine treatment under both normo- and hyperglycemic conditions. Gene set enrichment analysis (GSEA) showed that befotiamine increased peroxisomal lipid oxidation and organelle (mitochondrial) membrane function. In conclusion, benfotiamine increases mitochondrial glucose oxidation in myotubes and downregulates NOX4 expression. These findings may be of relevance to type 2 diabetes where reversal of reduced glucose oxidation and mitochondrial capacity is a desirable goal.


Benfotiamine Thiamine Myotubes Diabetes Hyperglycemia 



Nicotinamide adenine dinucleotide


Nicotinamide adenine dinucleotide phosphate


Nicotinamide adenine dinucleotide phosphate oxidase




Pentose phosphate pathway


Pyruvate dehydrogenase


Ketoglutarate dehydrogenase


Tricarboxylic acid


Intramyocellular triacylglycerol


Scintillation proximity assay






Growth factor midkine


Thiamine monophosphate


Thiamine diphosphate


Reduced folate carrier


Oxidative phosphorylation

Supplementary material

12263_2011_252_MOESM1_ESM.xls (61 kb)
Supplementary material 1 (XLS 61 kb)
12263_2011_252_MOESM2_ESM.xls (47 kb)
Supplementary material 2 (XLS 47 kb)
12263_2011_252_MOESM3_ESM.xlsx (1.6 mb)
Supplementary material 3 (XLSX 1588 kb)


  1. Aas V, Kase ET, Solberg R, Jensen J, Rustan AC (2004) Chronic hyperglycaemia promotes lipogenesis and triacylglycerol accumulation in human skeletal muscle cells. Diabetologia 47(8):1452–1461PubMedCrossRefGoogle Scholar
  2. Aas V, Hessvik NP, Wettergreen M, Hvammen AW, Hallen S, Thoresen GH, Rustan A (2011) Chronic hyperglycemia reduces substrate oxidation and impairs metabolic switching of human myotubes. Biochim Biophys Acta Mol Basis Dis 1812:94–105Google Scholar
  3. Alkhalaf A, Klooster A, van Oeveren W, Achenbach U, Kleefstra N, Slingerland RJ, Mijnhout GS, Bilo HJ, Gans RO, Navis GJ, Bakker SJ (2010) A double-blind, randomized, placebo-controlled clinical trial on benfotiamine treatment in patients with diabetic nephropathy. Diabetes Care 33(7):1598–1601PubMedCrossRefGoogle Scholar
  4. Babaei-Jadidi R, Karachalias N, Ahmed N, Battah S, Thornalley PJ (2003) Prevention of incipient diabetic nephropathy by high-dose thiamine and benfotiamine. Diabetes 52(8):2110–2120PubMedCrossRefGoogle Scholar
  5. Balakumar P, Rohilla A, Krishan P, Solairaj P, Thangathirupathi A (2010) The multifaceted therapeutic potential of benfotiamine. Pharmacol Res 61(6):482–488PubMedCrossRefGoogle Scholar
  6. Basuroy S, Bhattacharya S, Leffler CW, Parfenova H (2009) Nox4 NADPH oxidase mediates oxidative stress and apoptosis caused by TNF-alpha in cerebral vascular endothelial cells. Am J Phys Cell Physiol 296(3):C422–C432CrossRefGoogle Scholar
  7. Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Statist Soc B 57(1):289–300Google Scholar
  8. Bohn SK, Myhrstad MC, Thoresen M, Holden M, Karlsen A, Tunheim SH, Erlund I, Svendsen M, Seljeflot I, Moskaug JO, Duttaroy AK, Laake P, Arnesen H, Tonstad S, Collins A, Drevon CA, Blomhoff R (2010) Blood cell gene expression associated with cellular stress defense is modulated by antioxidant-rich food in a randomised controlled clinical trial of male smokers. BMC Med 8:54PubMedCrossRefGoogle Scholar
  9. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  10. Brazma A, Hingamp P, Quackenbush J et al (2001) Minimum information about a microarray experiment (MIAME)-toward standards for microarray data. Nat Genet 29(4):365–371Google Scholar
  11. Brinchmann-Hansen O, Dahl-Jorgensen K, Sandvik L, Hanssen KF (1992) Blood glucose concentrations and progression of diabetic retinopathy: the seven year results of the Oslo study. BMJ 304(6818):19–22PubMedCrossRefGoogle Scholar
  12. Brownlee M (2001) Biochemistry and molecular cell biology of diabetic complications. Nature 414(6865):813–820PubMedCrossRefGoogle Scholar
  13. Brownlee M (2005) The pathobiology of diabetic complications: a unifying mechanism. Diabetes 54(6):1615–1625PubMedCrossRefGoogle Scholar
  14. Chen K, Kirber MT, Xiao H, Yang Y, Keaney JF Jr (2008) Regulation of ROS signal transduction by NADPH oxidase 4 localization. J Cell Biol 181(7):1129–1139PubMedCrossRefGoogle Scholar
  15. Edwards LG, Adesida A, Thornalley PJ (1996) Inhibition of human leukaemia 60 cell growth by S-D-lactoylglutathione in vitro. Mediation by metabolism to N-D-lactoylcysteine and induction of apoptosis. Leukemia Res 20(1):17–26Google Scholar
  16. Gorin Y, Block K, Hernandez J, Bhandari B, Wagner B, Barnes JL, Abboud HE (2005) Nox4 NAD(P)H oxidase mediates hypertrophy and fibronectin expression in the diabetic kidney. J Biol Chem 280(47):39616–39626PubMedCrossRefGoogle Scholar
  17. Hammes HP, Du X, Edelstein D, Taguchi T, Matsumura T, Ju Q, Lin J, Bierhaus A, Nawroth P, Hannak D, Neumaier M, Bergfeld R, Giardino I, Brownlee M (2003) Benfotiamine blocks three major pathways of hyperglycemic damage and prevents experimental diabetic retinopathy. Nat Med 9(3):294–299PubMedCrossRefGoogle Scholar
  18. Hilbig R, Rahmann H (1998) Comparative autoradiographic investigations on the tissue distribution of benfotiamine versus thiamine in mice. Arzneimittelforschung 48(5):461–468PubMedGoogle Scholar
  19. Kosugi T, Yuzawa Y, Sato W, Kawai H, Matsuo S, Takei Y, Muramatsu T, Kadomatsu K (2006) Growth factor midkine is involved in the pathogenesis of diabetic nephropathy. Am J Pathol 168(1):9–19PubMedCrossRefGoogle Scholar
  20. Loew D (1996) Pharmacokinetics of thiamine derivatives especially of benfotiamine. Int J Clin Pharmacol Ther 34(2):47–50PubMedGoogle Scholar
  21. Miyata T, Inagi R, Nangaku M, Imasawa T, Sato M, Izuhara Y, Suzuki D, Yoshino A, Onogi H, Kimura M, Sugiyama S, Kurokawa K (2002) Overexpression of the serpin megsin induces progressive mesangial cell proliferation and expansion. J Clin Invest 109(5):585–593PubMedGoogle Scholar
  22. Ohtomo S, Nangaku M, Izuhara Y, Yamada N, Dan T, Mori T, Ito S, de Strihou C, Miyata T (2008) The role of megsin, a serine protease inhibitor, in diabetic mesangial matrix accumulation. Kidney Int 74(6):768–774PubMedCrossRefGoogle Scholar
  23. Pagel-Langenickel I, Bao J, Pang L, Sack MN (2010) The role of mitochondria in the pathophysiology of skeletal muscle insulin resistance. Endocrine Rev 31(1):25–51CrossRefGoogle Scholar
  24. Richter EA, Hansen BF, Hansen SA (1988) Glucose-induced insulin resistance of skeletal-muscle glucose transport and uptake. Biochem J 252(3):733–737PubMedGoogle Scholar
  25. Sartor MA, Tomlinson CR, Wesselkamper SC, Sivaganesan S, Leikauf GD, Medvedovic M (2006) Intensity-based hierarchical Bayes method improves testing for differentially expressed genes in microarray experiments. BMC Bioinformatics 7:538PubMedCrossRefGoogle Scholar
  26. Schmid U, Stopper H, Heidland A, Schupp N (2008) Benfotiamine exhibits direct antioxidative capacity and prevents induction of DNA damage in vitro. Diabetes Metab Res Rev 24(5):371–377PubMedCrossRefGoogle Scholar
  27. Schupp N, Dette EM, Schmid U, Bahner U, Winkler M, Heidland A, Stopper H (2008) Benfotiamine reduces genomic damage in peripheral lymphocytes of hemodialysis patients. Naunyn Schmiedebergs Arch Pharmacol 378(3):283–291PubMedCrossRefGoogle Scholar
  28. Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, Paulovich A, Pomeroy SL, Golub TR, Lander ES, Mesirov JP (2005) Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci 102(43):15545–15550PubMedCrossRefGoogle Scholar
  29. The 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(14):977–986CrossRefGoogle Scholar
  30. Thornalley PJ, Babaei-Jadidi R (2005) Prevention of microvascular complications of diabetes by high dose S-benzoylthiamine monophosphate (Benfotiamine): mechanism of thiamine delivery into cells. Diabetologia 48:(Suppl 1)Google Scholar
  31. Tilton RG, Baier LD, Harlow JE, Smith SR, Ostrow E, Williamson JR (1992) Diabetes-induced glomerular dysfunction: links to a more reduced cytosolic ratio of NADH/NAD+. Kidney Int 41(4):778–788PubMedCrossRefGoogle Scholar
  32. UK Prospective Diabetes Study (UKPDS) Group (1998) Intensive blood-glucose control with sulphonylureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes (UKPDS 33). Lancet 352(9131):837–853CrossRefGoogle Scholar
  33. Van den Enden MK, Nyengaard JR, Ostrow E, Burgan JH, Williamson JR (1995) Elevated glucose levels increase retinal glycolysis and sorbitol pathway metabolism. Implications for diabetic retinopathy. Invest Ophth Vis Sci 36(8):1675–1685Google Scholar
  34. Volvert ML, Seyen S, Piette M, Evrard B, Gangolf M, Plumier JC, Bettendorff L (2008) Benfotiamine, a synthetic S-acyl thiamine derivative, has different mechanisms of action and a different pharmacological profile than lipid-soluble thiamine disulfide derivatives. BMC Pharmacol 8:10PubMedCrossRefGoogle Scholar
  35. Wensaas AJ, Rustan AC, Lovstedt K, Kull B, Wikstrom S, Drevon CA, Hallen S (2007) Cell-based multiwell assays for the detection of substrate accumulation and oxidation. J Lipid Res 48(4):961–967PubMedCrossRefGoogle Scholar
  36. Xia L, Wang H, Goldberg HJ, Munk S, Fantus IG, Whiteside CI (2006) Mesangial cell NADPH oxidase upregulation in high glucose is protein kinase C dependent and required for collagen IV expression. Am J Physiol Renal Physiol 290(2):F345–F356PubMedCrossRefGoogle Scholar
  37. Xu M, Dai DZ, Dai Y (2009) Normalizing NADPH oxidase contributes to attenuating diabetic nephropathy by the dual endothelin receptor antagonist CPU0213 in rats. Am J Nephrol 29(3):252–256PubMedCrossRefGoogle Scholar
  38. Ziems M, Netzel M, Bitsch I (2000) Biokinetic parameters and metabolism of S-benzoylthiamine-O-monophosphate. Biofactors 11(1–2):109–110PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • D. A. Fraser
    • 1
  • N. P. Hessvik
    • 2
  • N. Nikolić
    • 2
  • V. Aas
    • 3
  • K. F. Hanssen
    • 4
  • S. K. Bøhn
    • 5
  • G. H. Thoresen
    • 2
  • A. C. Rustan
    • 2
  1. 1.Diabetes Research CentreOslo University HospitalOsloNorway
  2. 2.Department of Pharmaceutical Biosciences, School of PharmacyUniversity of OsloOsloNorway
  3. 3.Faculty of Health SciencesOslo University CollegeOsloNorway
  4. 4.Department of EndocrinologyOslo University HospitalOsloNorway
  5. 5.Department of Nutrition, Faculty of Medicine, Institute of Basic Medical SciencesUniversity of OsloOsloNorway

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