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Effect of Alcohol Administration on Mg(2+) Homeostasis in H9C2 Cells.

Nguyen H, Romani A - J Cardiovasc Dis Diagn (2014)

Bottom Line: This pathology contrasts the seemingly beneficial effect of small doses of alcohol on the cardiovascular system.EtOH-induced Mg(2+) extrusion was also prevented by the administration of di-thio-treitol (DTT) and n-acetyl-cysteine (NAC), two agents that prevent the negative effects of ROS formation and free radicals generation associated with EtOH metabolism by cyP4502E1.Taken together, our data indicate that Mg(2+) extrusion also occur in cardiac cells exposed to EtOH as a result of alcohol metabolism by cyP4502E1 and associated free radical formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology and Biophysics, School of Medicine, Case Western Reserve University, Cleveland, OH 44106, USA.

ABSTRACT

Alcoholic cardiomyopathy represents one of the main clinical complications in chronic alcoholics. This pathology contrasts the seemingly beneficial effect of small doses of alcohol on the cardiovascular system. Studies carried out in liver cells exposed acutely or chronically to varying doses of EtOH indicate that intrahepatic alcohol metabolism results in a major loss of cellular Mg(2+). To investigate whether EtOH administration also induced Mg(2+) extrusion in cardiac cells, H9C2 cells were exposed to varying doses of EtOH for short- or ling-term periods of time. The results indicate that H9C2 cells exposed to EtOH doses higher than 0.1% (v/v, or 15 mM) extruded Mg(2+) into the extracellular medium on a time- and dose-dependent manner. Consistent with the involvement of cyP4502E1 in metabolizing EtOH, administration of chloro-methiazole (CMZ) as an inhibitor of the cytochrome prevented EtOH-induced Mg(2+) loss to a large extent. EtOH-induced Mg(2+) extrusion was also prevented by the administration of di-thio-treitol (DTT) and n-acetyl-cysteine (NAC), two agents that prevent the negative effects of ROS formation and free radicals generation associated with EtOH metabolism by cyP4502E1. Taken together, our data indicate that Mg(2+) extrusion also occur in cardiac cells exposed to EtOH as a result of alcohol metabolism by cyP4502E1 and associated free radical formation. Interestingly, Mg(2+) extrusion only occurs at doses of EtOH higher than 0.1% administered for an extended period of time. The significance of Mg(2+) extrusion for the onset of alcoholic cardiomyopathy remains to be elucidated.

No MeSH data available.


Related in: MedlinePlus

Ethanol-induced Mg2+ extrusion in H9C2 cellsH9C2 cells, plated as indicated under Material and Methods, were stimulated by addition of varying doses of EtOH to the incubation medium. At the time points reported in the figure, aliquots of extracellular medium were withdrawn and Mg2+ content was assessed by AAS. Figure 1A reports a typical Mg2+ extrusion profile for H9C2 cells. Net Mg2+ extrusion is reported in Figure 1B. Data reported in Figure 1A and 1B are means ± S.E. of 5 different cell preparations, each tested in duplicate for all the experimental conditions. *Statistical significant (p<0.01) vs. corresponding time points in control sample and 0.01% stimulated cells.
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Figure 1: Ethanol-induced Mg2+ extrusion in H9C2 cellsH9C2 cells, plated as indicated under Material and Methods, were stimulated by addition of varying doses of EtOH to the incubation medium. At the time points reported in the figure, aliquots of extracellular medium were withdrawn and Mg2+ content was assessed by AAS. Figure 1A reports a typical Mg2+ extrusion profile for H9C2 cells. Net Mg2+ extrusion is reported in Figure 1B. Data reported in Figure 1A and 1B are means ± S.E. of 5 different cell preparations, each tested in duplicate for all the experimental conditions. *Statistical significant (p<0.01) vs. corresponding time points in control sample and 0.01% stimulated cells.

Mentions: As Figure 1 shows, H9C2 cells exposed to EtOH extruded Mg2+ across the cell membrane into the extracellular medium in a dose- and a time-dependent manner (Figure 1A). The Mg2+ extrusion was observed as net increase in the extracellular medium (Figure 1B), or as a decrease in total cellular Mg2+ content (Figure 2A). Exposure to EtOH for 24 h also resulted in a detectable Mg2+ loss (Figure 2B) that was slightly higher than that observed in cells exposed to EtOH for 60 min (Figure 2A).


Effect of Alcohol Administration on Mg(2+) Homeostasis in H9C2 Cells.

Nguyen H, Romani A - J Cardiovasc Dis Diagn (2014)

Ethanol-induced Mg2+ extrusion in H9C2 cellsH9C2 cells, plated as indicated under Material and Methods, were stimulated by addition of varying doses of EtOH to the incubation medium. At the time points reported in the figure, aliquots of extracellular medium were withdrawn and Mg2+ content was assessed by AAS. Figure 1A reports a typical Mg2+ extrusion profile for H9C2 cells. Net Mg2+ extrusion is reported in Figure 1B. Data reported in Figure 1A and 1B are means ± S.E. of 5 different cell preparations, each tested in duplicate for all the experimental conditions. *Statistical significant (p<0.01) vs. corresponding time points in control sample and 0.01% stimulated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362511&req=5

Figure 1: Ethanol-induced Mg2+ extrusion in H9C2 cellsH9C2 cells, plated as indicated under Material and Methods, were stimulated by addition of varying doses of EtOH to the incubation medium. At the time points reported in the figure, aliquots of extracellular medium were withdrawn and Mg2+ content was assessed by AAS. Figure 1A reports a typical Mg2+ extrusion profile for H9C2 cells. Net Mg2+ extrusion is reported in Figure 1B. Data reported in Figure 1A and 1B are means ± S.E. of 5 different cell preparations, each tested in duplicate for all the experimental conditions. *Statistical significant (p<0.01) vs. corresponding time points in control sample and 0.01% stimulated cells.
Mentions: As Figure 1 shows, H9C2 cells exposed to EtOH extruded Mg2+ across the cell membrane into the extracellular medium in a dose- and a time-dependent manner (Figure 1A). The Mg2+ extrusion was observed as net increase in the extracellular medium (Figure 1B), or as a decrease in total cellular Mg2+ content (Figure 2A). Exposure to EtOH for 24 h also resulted in a detectable Mg2+ loss (Figure 2B) that was slightly higher than that observed in cells exposed to EtOH for 60 min (Figure 2A).

Bottom Line: This pathology contrasts the seemingly beneficial effect of small doses of alcohol on the cardiovascular system.EtOH-induced Mg(2+) extrusion was also prevented by the administration of di-thio-treitol (DTT) and n-acetyl-cysteine (NAC), two agents that prevent the negative effects of ROS formation and free radicals generation associated with EtOH metabolism by cyP4502E1.Taken together, our data indicate that Mg(2+) extrusion also occur in cardiac cells exposed to EtOH as a result of alcohol metabolism by cyP4502E1 and associated free radical formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology and Biophysics, School of Medicine, Case Western Reserve University, Cleveland, OH 44106, USA.

ABSTRACT

Alcoholic cardiomyopathy represents one of the main clinical complications in chronic alcoholics. This pathology contrasts the seemingly beneficial effect of small doses of alcohol on the cardiovascular system. Studies carried out in liver cells exposed acutely or chronically to varying doses of EtOH indicate that intrahepatic alcohol metabolism results in a major loss of cellular Mg(2+). To investigate whether EtOH administration also induced Mg(2+) extrusion in cardiac cells, H9C2 cells were exposed to varying doses of EtOH for short- or ling-term periods of time. The results indicate that H9C2 cells exposed to EtOH doses higher than 0.1% (v/v, or 15 mM) extruded Mg(2+) into the extracellular medium on a time- and dose-dependent manner. Consistent with the involvement of cyP4502E1 in metabolizing EtOH, administration of chloro-methiazole (CMZ) as an inhibitor of the cytochrome prevented EtOH-induced Mg(2+) loss to a large extent. EtOH-induced Mg(2+) extrusion was also prevented by the administration of di-thio-treitol (DTT) and n-acetyl-cysteine (NAC), two agents that prevent the negative effects of ROS formation and free radicals generation associated with EtOH metabolism by cyP4502E1. Taken together, our data indicate that Mg(2+) extrusion also occur in cardiac cells exposed to EtOH as a result of alcohol metabolism by cyP4502E1 and associated free radical formation. Interestingly, Mg(2+) extrusion only occurs at doses of EtOH higher than 0.1% administered for an extended period of time. The significance of Mg(2+) extrusion for the onset of alcoholic cardiomyopathy remains to be elucidated.

No MeSH data available.


Related in: MedlinePlus