Suppression of the accumulation of triosephosphates and increased formation of methylglyoxal in human red blood cells during hyperglycaemia by thiamine in vitro.
The accumulation of triosephosphates and the increased formation of the potent glycating agent methylglyoxal in intracellular hyperglycaemia are implicated in the development of diabetic complications. A strategy to counter this is to stimulate the anaerobic pentosephosphate pathway of glycolysis by maximizing transketolase activity by thiamine supplementation, with the consequent consumption of glyceraldehyde-3-phosphate and increased formation of ribose-5-phosphate. To assess the effect of thiamine supplementation on the accumulation of triosephosphates and methylglyoxal formation in cellular hyperglycaemia, we incubated human red blood cell suspensions (50% v/v) in short-term culture with 5 mM glucose and 50 mM glucose in Krebs-Ringer phosphate buffer at 37 degrees C as models of cellular metabolism under normoglycaemic and hyperglycaemic conditions. In hyperglycaemia, there is a characteristic increase in the concentration of the triosephosphate pool of glycolytic intermediates and a consequent increase in the concentration and metabolic flux of the formation of methylglyoxal. The addition of thiamine (50-500 microM) increased the activity of transketolase, decreased the concentration of the triosephosphate pool, decreased the concentration and metabolic flux of the formation of methylglyoxal, and increased the concentration of total sedoheptulose-7-phosphate and ribose-5-phosphate. Biochemical changes implicated in the development of diabetic complications were thereby prevented. This provides a biochemical basis for high dose thiamine therapy for the prevention of diabetic complications.
Thornalley
P J
PJ
Department of Biological Sciences, University of Essex, Central Campus, Wivenhoe Park, Colchester CO4 3SQ, Essex, UK. thorp@essex.ac.uk
Jahan
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D003847:Deoxyglucose / Q000031:analogs & derivatives / Q000378:metabolism; D048909:Diabetes Complications; D003920:Diabetes Mellitus / Q000097:blood / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects* / Q000378:metabolism; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006019:Glycolysis / Q000187:drug effects*; D006801:Humans; D006943:Hyperglycemia / Q000097:blood* / Q000150:complications / Q000188:drug therapy / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D010428:Pentosephosphates / Q000378:metabolism; D011765:Pyruvaldehyde / Q000378:metabolism*; D019289:Pyruvic Acid / Q000378:metabolism; D013403:Sugar Phosphates / Q000378:metabolism*; D013831:Thiamine / Q000494:pharmacology* / Q000627:therapeutic use; D014174:Transketolase / Q000378:metabolism; D014306:Trioses / Q000378:metabolism
D010428:Pentosephosphates; D013403:Sugar Phosphates; D014306:Trioses; D019344:Lactic Acid; D011765:Pyruvaldehyde; D019289:Pyruvic Acid; D003847:Deoxyglucose; D014174:Transketolase; C016350:3-deoxyglucosone; D005947:Glucose; D013831:Thiamine