An environment of high glucose concentration stimulates the synthesis of extracellular matrix (ECM) in mesangial cell (MC) cultures. This may result from a similar increase in intracellular glucose concentration. We theorized that increased uptake, rather than glucose concentration per se is the major determinant of exaggerated ECM formation. To test this, we compared the effects of 35 mM glucose on ECM synthesis in normal MCs with those of 8 mM glucose in the same cells overexpressing the glucose transporter GLUT1 (MCGT1). Increasing medium glucose from 8 to 35 mM caused normal MCs to increase total collagen synthesis and catabolism, with a net 81-90% increase in accumulation. MCs transduced with the human GLUT] gene (MCGT1) grown in 8 mM glucose had a 10-fold greater GLUT1 protein expression and a 1.9, 2.1, and 2.5-fold increase in cell myo-inositol, lactate production, and cell sorbitol content, respectively, as compared to control MCs transduced with bacterial f8-galactosidase (MCLacZ). MCGT1 also demonstrated increased glucose uptake (5-fold) and increased net utilization (43-fold), and greater synthesis of individual ECM components than MCLacZ. In addition, total collagen synthesis and catabolism were also enhanced with a net collagen accumulation 111-118% greater than controls. Thus, glucose transport activity is an important modulator of ECM formation by MCs; the presence of high extracellular glucose concentrations is not necessarily required for the stimulation of matrix synthesis. (J.

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Glucose transporter protein type 1 (GLUT1) is a major glucose transporter of the fertilized egg and preimplantation embryo. Haploinsufficiency for GLUT1 causes the GLUT1 deficiency syndrome in humans, however the embryo appears unaffected. Therefore, here we produced heterozygous GLUT1 knockout murine embryonic stem cells (GT1؉/؊) to study the role of GLUT1 deficiency in their growth, glucose metabolism, and survival in response to hypoxic stress. GT1(؊/؊) cells were determined to be nonviable. Both the GLUT1 and GLUT3 high-affinity, facilitative glucose transporters were expressed in GT1(؉/؉) and GT1(؉/؊) embryonic stem cells. GT1(؉/؊) demonstrated 49 ؎ 4% reduction of GLUT1 mRNA. This induced a posttranscriptional, GLUT1 compensatory response resulting in 24 ؎ 4% reduction of GLUT1 protein. GLUT3 was unchanged. GLUT8 and GLUT12 were also expressed and unchanged in GT1(؉/؊). Stimulation of glycolysis by azide inhibition of oxidative phosphorylation was impaired by 44% in GT1(؉/؊), with impaired up-regulation of GLUT1 protein. Hypoxia for up to 4 hours led to 201% more apoptosis in GT1(؉/؊) than in GT1(؉/؉) controls. Caspase-3 activity was 76% higher in GT1(؉/؊) versus GT1(؉/؉) at 2 hours. Heterozygous knockout of GLUT1 led to a partial GLUT1 compensatory response protecting nonstressed cells. However, inhibition of oxidative phosphorylation and hypoxia both exposed their increased susceptibility to these stresses.

Metformin (MTF) is one of the most common oral agents used to treat diabetes mellitus. Intoxication is associated with lactic acidosis and has significant clinical consequences. We report 12 cases requiring dialytic intervention. Twelve patients were analyzed from 2005 to 2010; 10 of these patients were treated with dialysis. Conventional hemodialysis (HD) and continuous veno-venous hemodialysis treatments with bicarbonate dialysis were used, and the results were presented as mean and standard deviation. The results are as follows: 33% of the patients were male, hospital stay was 9.3 (± 12) days, average MTF dose 1.7 g/day, mortality was 25%. Baseline glomerular filtration rate for these patients was 51.5 mL/min, with an average age of 64 (± 11) years. On presentation, all had acute kidney injury with blood urea nitrogen/creatinine 75 (± 30)/8.1 (± 3.7) mg/dL, lactic acid 12.4 (± 8.1) mmol/L, pH 7.04 (± 0.19), bicarbonate 7.2 (± 4.5) mmol/L. Metformin level was 25 (± 17) µg/mL; anion gap was 28 (± 9), and serum potassium was 5.4 (± 1.3) mEq/L. Seventy percent of patients were treated with conventional HD. Patients required 4 (± 5) dialysis treatments at blood flow QB 330 (± 53), dialysis flow QD 571 (± 111) for 305 (± 122) minutes. Postdialysis, the acidosis parameters improved: bicarbonate 19.2 (± 4.1) mmol/L, lactic acid 6 (± 4) mmol/L and MTF levels decreased 8.9 (± 5.7) µg/mL. Metformin percentage removal was calculated to be 60% (± 24). No difference was found between HD and continous veno-venous hemodialysis. The only difference between survivors was the age 53 (± 7) vs. 78 (± 10) (P < 0.05). Metformin toxicity is a serious clinical condition and causes severe lactic acidosis and significant mortality. Hemodialysis is an efficient method to treat MTF intoxication and correct the metabolic abnormalities.