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Adenosine 5'-triphosphate stimulates the increase of TGF-beta1 in rat mesangial cells under high-glucose conditions via reactive oxygen species and ERK1/2.

Qu LP, Xue H, Yuan P, Zhou L, Yao T, Huang Y, Lu LM - Acta Pharmacol. Sin. (2009)

Bottom Line: The effects of high glucose on TGF-beta1 mRNA were markedly inhibited by suramin, DPI or PD98059.Our results suggest that a high concentration of glucose increases the extracellular levels of ATP in mesangial cells within a short time-frame.ATP, in turn, activates ERK1/2, an effect which is at least partially dependent on ROS, which results in the upregulation of TGF-beta1.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology and Pathophysiology, Shanghai Medical College, Fudan University, Shanghai, China.

ABSTRACT

Aim: To investigate the role of adenosine 5'-triphosphate (ATP)-induced generation of reactive oxygen species (ROS) and phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in the production of transforming growth factor-beta1 (TGF-beta1) in cultured rat glomerular mesangial cells under high-glucose conditions.

Methods: Subconfluent glomerular mesangial cells were serum-starved for 24 h and pretreated with suramin, diphenylenechloride iodonium (DPI) or PD98059 followed by stimulation with a high concentration of glucose (30 mmol/L D-glucose) or ATP (300 micromol/L). Extracellular and total ATP and ROS production were detected using commercially available kits. Phosphorylation of ERK1/2 was evaluated by Western blot. TGF-beta1 mRNA expression was examined by real-time PCR.

Results: Suramin had a dose-dependent inhibitory effect on the generation of ROS induced by high glucose. Extracellular ATP production by mesangial cells increased markedly after a 2-h incubation with high glucose. ROS production was upregulated in mesangial cells after 5 min incubation with 300 micromol/L ATP and was sustained for 120 min. ERK1/2 was significantly activated after 5 min incubation of mesangial cells with ATP, this activation was partially inhibited by DPI. The effects of high glucose on TGF-beta1 mRNA were markedly inhibited by suramin, DPI or PD98059.

Conclusion: Our results suggest that a high concentration of glucose increases the extracellular levels of ATP in mesangial cells within a short time-frame. ATP, in turn, activates ERK1/2, an effect which is at least partially dependent on ROS, which results in the upregulation of TGF-beta1.

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Related in: MedlinePlus

Effect of suramin on the generation of ROS induced by high glucose in mesangial cells. Rat mesangial cells were cultured under high glucose (30 mmol/L D-glucose) conditions for 2 h. In order to explore the involvement of extracellular ATP in the generation of ROS, the cells were pretreated with suramin, a nonspecific P2 receptor antagonist, at the indicated concentrations for 30 min. n=4–6. Mean±SEM. aP>0.05 vs control; bP<0.05 vs control; eP<0.05 vs HG (high glucose).
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fig1: Effect of suramin on the generation of ROS induced by high glucose in mesangial cells. Rat mesangial cells were cultured under high glucose (30 mmol/L D-glucose) conditions for 2 h. In order to explore the involvement of extracellular ATP in the generation of ROS, the cells were pretreated with suramin, a nonspecific P2 receptor antagonist, at the indicated concentrations for 30 min. n=4–6. Mean±SEM. aP>0.05 vs control; bP<0.05 vs control; eP<0.05 vs HG (high glucose).

Mentions: Suramin exhibited a dose-dependent inhibitory effect on the increased generation of ROS induced by high glucose for 2 h in rat mesangial cells; suramin at 100 μmol/L had an obvious effect on the diminishment of ROS generation (P<0.05 vs HG, high glucose; Figure 1).


Adenosine 5'-triphosphate stimulates the increase of TGF-beta1 in rat mesangial cells under high-glucose conditions via reactive oxygen species and ERK1/2.

Qu LP, Xue H, Yuan P, Zhou L, Yao T, Huang Y, Lu LM - Acta Pharmacol. Sin. (2009)

Effect of suramin on the generation of ROS induced by high glucose in mesangial cells. Rat mesangial cells were cultured under high glucose (30 mmol/L D-glucose) conditions for 2 h. In order to explore the involvement of extracellular ATP in the generation of ROS, the cells were pretreated with suramin, a nonspecific P2 receptor antagonist, at the indicated concentrations for 30 min. n=4–6. Mean±SEM. aP>0.05 vs control; bP<0.05 vs control; eP<0.05 vs HG (high glucose).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4007501&req=5

fig1: Effect of suramin on the generation of ROS induced by high glucose in mesangial cells. Rat mesangial cells were cultured under high glucose (30 mmol/L D-glucose) conditions for 2 h. In order to explore the involvement of extracellular ATP in the generation of ROS, the cells were pretreated with suramin, a nonspecific P2 receptor antagonist, at the indicated concentrations for 30 min. n=4–6. Mean±SEM. aP>0.05 vs control; bP<0.05 vs control; eP<0.05 vs HG (high glucose).
Mentions: Suramin exhibited a dose-dependent inhibitory effect on the increased generation of ROS induced by high glucose for 2 h in rat mesangial cells; suramin at 100 μmol/L had an obvious effect on the diminishment of ROS generation (P<0.05 vs HG, high glucose; Figure 1).

Bottom Line: The effects of high glucose on TGF-beta1 mRNA were markedly inhibited by suramin, DPI or PD98059.Our results suggest that a high concentration of glucose increases the extracellular levels of ATP in mesangial cells within a short time-frame.ATP, in turn, activates ERK1/2, an effect which is at least partially dependent on ROS, which results in the upregulation of TGF-beta1.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology and Pathophysiology, Shanghai Medical College, Fudan University, Shanghai, China.

ABSTRACT

Aim: To investigate the role of adenosine 5'-triphosphate (ATP)-induced generation of reactive oxygen species (ROS) and phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in the production of transforming growth factor-beta1 (TGF-beta1) in cultured rat glomerular mesangial cells under high-glucose conditions.

Methods: Subconfluent glomerular mesangial cells were serum-starved for 24 h and pretreated with suramin, diphenylenechloride iodonium (DPI) or PD98059 followed by stimulation with a high concentration of glucose (30 mmol/L D-glucose) or ATP (300 micromol/L). Extracellular and total ATP and ROS production were detected using commercially available kits. Phosphorylation of ERK1/2 was evaluated by Western blot. TGF-beta1 mRNA expression was examined by real-time PCR.

Results: Suramin had a dose-dependent inhibitory effect on the generation of ROS induced by high glucose. Extracellular ATP production by mesangial cells increased markedly after a 2-h incubation with high glucose. ROS production was upregulated in mesangial cells after 5 min incubation with 300 micromol/L ATP and was sustained for 120 min. ERK1/2 was significantly activated after 5 min incubation of mesangial cells with ATP, this activation was partially inhibited by DPI. The effects of high glucose on TGF-beta1 mRNA were markedly inhibited by suramin, DPI or PD98059.

Conclusion: Our results suggest that a high concentration of glucose increases the extracellular levels of ATP in mesangial cells within a short time-frame. ATP, in turn, activates ERK1/2, an effect which is at least partially dependent on ROS, which results in the upregulation of TGF-beta1.

Show MeSH
Related in: MedlinePlus