To evaluate the role of glucose transporter- l (GLUT1) in the glucose uptake of glomerular mesangial cells. Methods. Cultured C57/SJL mouse mesangial cells were used in the study. The expression of GLUT1 mRNA was dete...To evaluate the role of glucose transporter- l (GLUT1) in the glucose uptake of glomerular mesangial cells. Methods. Cultured C57/SJL mouse mesangial cells were used in the study. The expression of GLUT1 mRNA was detected by RT- PCR. The expression of GLUT1 protein was detected by immunofluorescence and flow cytometry. The uptake of glucose and its kinetics were determined by 2- deoxy- [3H]- D- glucose uptake. Results. Both GLUT1 mRNA and protein were found in mouse glomerular mesangial cells. 2- deoxy- D- glucose uptake and kinetics assay showed that this glucose transporter had high affinity for glucose and the glucose uptake specificity was further confirmed by phloretin. Conclusion. Functional GLUT1 did present in mouse mesangial cells cultured in vitro and it might be the predominant transporter mediated the uptake of glucose into mesangial cells.展开更多
Objective To explore the effect of rhein on the regulation of glucose transporter 1 (GLUT1) overexpression and the possible molecular mechanism that rhein antagonize the effect of transforming growth factor β1 (TGF ...Objective To explore the effect of rhein on the regulation of glucose transporter 1 (GLUT1) overexpression and the possible molecular mechanism that rhein antagonize the effect of transforming growth factor β1 (TGF β1) in glomerular mesangial cells Methods Cultured mouse mesangial cells were used The expression of GLUT1 mRNA was detected by Northern blotting; the ability of glucose uptake was determined by 2 deoxy [ 3H] D glucose uptake assay Results Rhein had no effect on glucose uptake in mesangial cells cultured in normal glucose concentration TGF β1 could upregulate the expression of GLUT1 mRNA and glucose uptake in mesangial cells This effect was markedly attenuated by the addition of rhein in a dose dependent manner Conclusions TGF β1 could upregulate the expression of GLUT1 mRNA and glucose uptake in mesangial cells, resulting in excessive glucose consumption and extracellular matrix production in diabetic nephropathy Rhein antagonized the effect of TGF β1 in mesangial cells, so it might be a hopeful remedy for the treatment of patients with diabetic展开更多
Objective To study the effects of high glucose and transforming growth factor β1 (TGF β1) on the expression and function of glucose transporter 1 (GLUT1) in mouse mesangial cells Methods Cultured mouse mesangial cel...Objective To study the effects of high glucose and transforming growth factor β1 (TGF β1) on the expression and function of glucose transporter 1 (GLUT1) in mouse mesangial cells Methods Cultured mouse mesangial cells were used The expression of GLUT1 mRNA was detected by Northern Blot; glucose uptake and its kinetics were determined with a 2 Deoxy [ 3H] D glucose uptake assay Results Mesangial cells exposed to enriched glucose medium (20?mmol/L) for 72 hours demonstrated a decrease in both GLUT1 mRNA and V max for uptake of the glucose analog, 2 deoxy D glucose (2DOG), as compared to mesangial cells cultured in physiologic glucose concentrations(5 5?mmol/L) In contrast, hypertonic mannitol had no effect on GLUT1 mRNA levels TGF β1 treatment for 10 hours stimulated 2DOG uptake, both in 5 5?mmol/L and 20?mmol/L glucose medium, by approximately 4 28 fold in a dose dependent manner (2?ng/ml maximum) Kinetic analysis of 2DOG uptake revealed an increase in V max and a decrease in K m in the presence of TGF β1 TGF β1 also up regulated the expression of GLUT1 mRNA in mesangial cells The addition of anti TGF β neutralizing antibody (30?μg/ml) in mesangial cells cultured in enriched glucose medium (20?mmol/L) led to a 40% decrease in 2DOG uptake Conclusions The expression of GLUT1 can be suppressed by exposure of mesangial cells to high glucose medium, which may serve as a protective mechanism against possible adverse effects of excessive glucose flux into cells TGF β1 stimulates glucose uptake by enhancing the expression and function of GLUT1 in mesangial cells This effect is independent of the glucose milieu in the cultured展开更多
基金This work was supported by the National Natural Science Foundation of China (No.39870288)
文摘To evaluate the role of glucose transporter- l (GLUT1) in the glucose uptake of glomerular mesangial cells. Methods. Cultured C57/SJL mouse mesangial cells were used in the study. The expression of GLUT1 mRNA was detected by RT- PCR. The expression of GLUT1 protein was detected by immunofluorescence and flow cytometry. The uptake of glucose and its kinetics were determined by 2- deoxy- [3H]- D- glucose uptake. Results. Both GLUT1 mRNA and protein were found in mouse glomerular mesangial cells. 2- deoxy- D- glucose uptake and kinetics assay showed that this glucose transporter had high affinity for glucose and the glucose uptake specificity was further confirmed by phloretin. Conclusion. Functional GLUT1 did present in mouse mesangial cells cultured in vitro and it might be the predominant transporter mediated the uptake of glucose into mesangial cells.
文摘Objective To explore the effect of rhein on the regulation of glucose transporter 1 (GLUT1) overexpression and the possible molecular mechanism that rhein antagonize the effect of transforming growth factor β1 (TGF β1) in glomerular mesangial cells Methods Cultured mouse mesangial cells were used The expression of GLUT1 mRNA was detected by Northern blotting; the ability of glucose uptake was determined by 2 deoxy [ 3H] D glucose uptake assay Results Rhein had no effect on glucose uptake in mesangial cells cultured in normal glucose concentration TGF β1 could upregulate the expression of GLUT1 mRNA and glucose uptake in mesangial cells This effect was markedly attenuated by the addition of rhein in a dose dependent manner Conclusions TGF β1 could upregulate the expression of GLUT1 mRNA and glucose uptake in mesangial cells, resulting in excessive glucose consumption and extracellular matrix production in diabetic nephropathy Rhein antagonized the effect of TGF β1 in mesangial cells, so it might be a hopeful remedy for the treatment of patients with diabetic
文摘Objective To study the effects of high glucose and transforming growth factor β1 (TGF β1) on the expression and function of glucose transporter 1 (GLUT1) in mouse mesangial cells Methods Cultured mouse mesangial cells were used The expression of GLUT1 mRNA was detected by Northern Blot; glucose uptake and its kinetics were determined with a 2 Deoxy [ 3H] D glucose uptake assay Results Mesangial cells exposed to enriched glucose medium (20?mmol/L) for 72 hours demonstrated a decrease in both GLUT1 mRNA and V max for uptake of the glucose analog, 2 deoxy D glucose (2DOG), as compared to mesangial cells cultured in physiologic glucose concentrations(5 5?mmol/L) In contrast, hypertonic mannitol had no effect on GLUT1 mRNA levels TGF β1 treatment for 10 hours stimulated 2DOG uptake, both in 5 5?mmol/L and 20?mmol/L glucose medium, by approximately 4 28 fold in a dose dependent manner (2?ng/ml maximum) Kinetic analysis of 2DOG uptake revealed an increase in V max and a decrease in K m in the presence of TGF β1 TGF β1 also up regulated the expression of GLUT1 mRNA in mesangial cells The addition of anti TGF β neutralizing antibody (30?μg/ml) in mesangial cells cultured in enriched glucose medium (20?mmol/L) led to a 40% decrease in 2DOG uptake Conclusions The expression of GLUT1 can be suppressed by exposure of mesangial cells to high glucose medium, which may serve as a protective mechanism against possible adverse effects of excessive glucose flux into cells TGF β1 stimulates glucose uptake by enhancing the expression and function of GLUT1 in mesangial cells This effect is independent of the glucose milieu in the cultured
