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Role of TRPM7 channels in hyperglycemia-mediated injury of vascular endothelial cells.

Sun H, Leng T, Zeng Z, Gao X, Inoue K, Xiong ZG - PLoS ONE (2013)

Bottom Line: Human umbilical vein endothelial cells (HUVECs) were incubated in the presence or absence of high concentrations of D-glucose (HG) for 72 h.In contrast to D-glucose, exposure of HUVECs to high concentrations of L-glucose had no effect.HG increased reactive oxygen species (ROS) generation, cytotoxicity and decreased endothelial nitric oxide synthase protein expression, which could be attenuated by knockdown of TRPM7 with TRPM7 siRNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China ; Neuroscience Institute, Morehouse School of Medicine, Atlanta, Georgia, United States of America.

ABSTRACT
This study investigated the change of transient receptor potential melastatin 7 (TRPM7) expression by high glucose and its role in hyperglycemia induced injury of vascular endothelial cells. Human umbilical vein endothelial cells (HUVECs) were incubated in the presence or absence of high concentrations of D-glucose (HG) for 72 h. RT-PCR, Real-time PCR, Western blotting, Immunofluorescence staining and whole-cell patch-clamp recordings showed that TRPM7 mRNA, TRPM7 protein expression and TRPM7-like currents were increased in HUVECs following exposure to HG. In contrast to D-glucose, exposure of HUVECs to high concentrations of L-glucose had no effect. HG increased reactive oxygen species (ROS) generation, cytotoxicity and decreased endothelial nitric oxide synthase protein expression, which could be attenuated by knockdown of TRPM7 with TRPM7 siRNA. The protective effect of silencing TRPM7 against HG induced endothelial injury was abolished by U0126, an inhibitor of the extracellular signal-regulated kinase signaling pathway. These observations suggest that TRPM7 channels play an important role in hyperglycemia-induced injury of vascular endothelial cells.

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Effect of HG on TRPM7 protein expression in HUVECs.(A) Representative immunoblots showing TRPM7 protein expression in HUVECs with or without HG (30 mM) for 72h. (B) The corresponding bar graphs showing relative expression of TRPM7 protein normalized to beta-actin. (C) Representative TRPM7-like currents recorded in HUVECs cultured in control or HG (30 mM) for 72h. (D) TRPM7-like current density. **p<0.01 vs. HG; *p<0.05 vs. control. n=4 for immunoblotting and 17-18 cells patched for current recording.
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pone-0079540-g001: Effect of HG on TRPM7 protein expression in HUVECs.(A) Representative immunoblots showing TRPM7 protein expression in HUVECs with or without HG (30 mM) for 72h. (B) The corresponding bar graphs showing relative expression of TRPM7 protein normalized to beta-actin. (C) Representative TRPM7-like currents recorded in HUVECs cultured in control or HG (30 mM) for 72h. (D) TRPM7-like current density. **p<0.01 vs. HG; *p<0.05 vs. control. n=4 for immunoblotting and 17-18 cells patched for current recording.

Mentions: Data are expressed as the mean ± SEM, and analyzed using one-way ANOVA with or without post-hoc multiple comparison tests. Data in Figure 1D and Figure S3 were analyzed by Student’s t-test. A p value of <0.05 was considered significant.


Role of TRPM7 channels in hyperglycemia-mediated injury of vascular endothelial cells.

Sun H, Leng T, Zeng Z, Gao X, Inoue K, Xiong ZG - PLoS ONE (2013)

Effect of HG on TRPM7 protein expression in HUVECs.(A) Representative immunoblots showing TRPM7 protein expression in HUVECs with or without HG (30 mM) for 72h. (B) The corresponding bar graphs showing relative expression of TRPM7 protein normalized to beta-actin. (C) Representative TRPM7-like currents recorded in HUVECs cultured in control or HG (30 mM) for 72h. (D) TRPM7-like current density. **p<0.01 vs. HG; *p<0.05 vs. control. n=4 for immunoblotting and 17-18 cells patched for current recording.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815131&req=5

pone-0079540-g001: Effect of HG on TRPM7 protein expression in HUVECs.(A) Representative immunoblots showing TRPM7 protein expression in HUVECs with or without HG (30 mM) for 72h. (B) The corresponding bar graphs showing relative expression of TRPM7 protein normalized to beta-actin. (C) Representative TRPM7-like currents recorded in HUVECs cultured in control or HG (30 mM) for 72h. (D) TRPM7-like current density. **p<0.01 vs. HG; *p<0.05 vs. control. n=4 for immunoblotting and 17-18 cells patched for current recording.
Mentions: Data are expressed as the mean ± SEM, and analyzed using one-way ANOVA with or without post-hoc multiple comparison tests. Data in Figure 1D and Figure S3 were analyzed by Student’s t-test. A p value of <0.05 was considered significant.

Bottom Line: Human umbilical vein endothelial cells (HUVECs) were incubated in the presence or absence of high concentrations of D-glucose (HG) for 72 h.In contrast to D-glucose, exposure of HUVECs to high concentrations of L-glucose had no effect.HG increased reactive oxygen species (ROS) generation, cytotoxicity and decreased endothelial nitric oxide synthase protein expression, which could be attenuated by knockdown of TRPM7 with TRPM7 siRNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China ; Neuroscience Institute, Morehouse School of Medicine, Atlanta, Georgia, United States of America.

ABSTRACT
This study investigated the change of transient receptor potential melastatin 7 (TRPM7) expression by high glucose and its role in hyperglycemia induced injury of vascular endothelial cells. Human umbilical vein endothelial cells (HUVECs) were incubated in the presence or absence of high concentrations of D-glucose (HG) for 72 h. RT-PCR, Real-time PCR, Western blotting, Immunofluorescence staining and whole-cell patch-clamp recordings showed that TRPM7 mRNA, TRPM7 protein expression and TRPM7-like currents were increased in HUVECs following exposure to HG. In contrast to D-glucose, exposure of HUVECs to high concentrations of L-glucose had no effect. HG increased reactive oxygen species (ROS) generation, cytotoxicity and decreased endothelial nitric oxide synthase protein expression, which could be attenuated by knockdown of TRPM7 with TRPM7 siRNA. The protective effect of silencing TRPM7 against HG induced endothelial injury was abolished by U0126, an inhibitor of the extracellular signal-regulated kinase signaling pathway. These observations suggest that TRPM7 channels play an important role in hyperglycemia-induced injury of vascular endothelial cells.

Show MeSH
Related in: MedlinePlus