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Diminution of Oxidative Damage to Human Erythrocytes and Lymphocytes by Creatine: Possible Role of Creatine in Blood.

Qasim N, Mahmood R - PLoS ONE (2015)

Bottom Line: Recent reports have shown that Cr displays antioxidant activity which could explain its beneficial cellular effects.This was accompanied by decrease in glutathione levels.Creatinine, a by-product of Cr metabolism, was also shown to exert protective effects, although it was slightly less effective than Cr.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.

ABSTRACT
Creatine (Cr) is naturally produced in the body and stored in muscles where it is involved in energy generation. It is widely used, especially by athletes, as a staple supplement for improving physical performance. Recent reports have shown that Cr displays antioxidant activity which could explain its beneficial cellular effects. We have evaluated the ability of Cr to protect human erythrocytes and lymphocytes against oxidative damage. Erythrocytes were challenged with model oxidants, 2, 2'-azobis(2-amidinopropane) dihydrochloride (AAPH) and hydrogen peroxide (H2O2) in the presence and absence of Cr. Incubation of erythrocytes with oxidant alone increased hemolysis, methemoglobin levels, lipid peroxidation and protein carbonyl content. This was accompanied by decrease in glutathione levels. Antioxidant enzymes and antioxidant power of the cell were compromised while the activity of membrane bound enzyme was lowered. This suggests induction of oxidative stress in erythrocytes by AAPH and H2O2. However, Cr protected the erythrocytes by ameliorating the AAPH and H2O2 induced changes in these parameters. This protective effect was confirmed by electron microscopic analysis which showed that oxidant-induced cell damage was attenuated by Cr. No cellular alterations were induced by Cr alone even at 20 mM, the highest concentration used. Creatinine, a by-product of Cr metabolism, was also shown to exert protective effects, although it was slightly less effective than Cr. Human lymphocytes were similarly treated with H2O2 in absence and presence of different concentrations of Cr. Lymphocytes incubated with oxidant alone had alterations in various biochemical and antioxidant parameters including decrease in cell viability and induction of DNA damage. The presence of Cr attenuated all these H2O2-induced changes in lymphocytes. Thus, Cr can function as a blood antioxidant, protecting cells from oxidative damage, genotoxicity and can potentially increase their lifespan.

No MeSH data available.


Related in: MedlinePlus

Effect of creatine on AAPH/ H2O2-induced changes in ferric reducing ability of cells.Erythrocytes were treated with the indicated concentrations of H2O2/AAPH, in presence and absence of Cr. FRAP assay was done in hemolysates. Results are mean values ± SEM from six independent experiments using blood from different donors. *Significantly different from control (p<0.05).
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pone.0141975.g005: Effect of creatine on AAPH/ H2O2-induced changes in ferric reducing ability of cells.Erythrocytes were treated with the indicated concentrations of H2O2/AAPH, in presence and absence of Cr. FRAP assay was done in hemolysates. Results are mean values ± SEM from six independent experiments using blood from different donors. *Significantly different from control (p<0.05).

Mentions: Antioxidant power of the cell was determined from the ability to quench free radicals and reduce metal ions by employing the widely used DPPH and FRAP assays, respectively. The metal reducing activity was determined from the ability of hemolysates to reduce ferric ions to ferrous form using the FRAP assay. The treatment of erythrocytes with either oxidant alone lowered the ability of erythrocytes to reduce ferric ions but it was greatly restored by Cr (Fig 5). The DPPH radical scavenging activity of lysates was also lowered by treatment with oxidants alone but recovered when Cr was also present in the incubation medium (Fig 6).


Diminution of Oxidative Damage to Human Erythrocytes and Lymphocytes by Creatine: Possible Role of Creatine in Blood.

Qasim N, Mahmood R - PLoS ONE (2015)

Effect of creatine on AAPH/ H2O2-induced changes in ferric reducing ability of cells.Erythrocytes were treated with the indicated concentrations of H2O2/AAPH, in presence and absence of Cr. FRAP assay was done in hemolysates. Results are mean values ± SEM from six independent experiments using blood from different donors. *Significantly different from control (p<0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4640867&req=5

pone.0141975.g005: Effect of creatine on AAPH/ H2O2-induced changes in ferric reducing ability of cells.Erythrocytes were treated with the indicated concentrations of H2O2/AAPH, in presence and absence of Cr. FRAP assay was done in hemolysates. Results are mean values ± SEM from six independent experiments using blood from different donors. *Significantly different from control (p<0.05).
Mentions: Antioxidant power of the cell was determined from the ability to quench free radicals and reduce metal ions by employing the widely used DPPH and FRAP assays, respectively. The metal reducing activity was determined from the ability of hemolysates to reduce ferric ions to ferrous form using the FRAP assay. The treatment of erythrocytes with either oxidant alone lowered the ability of erythrocytes to reduce ferric ions but it was greatly restored by Cr (Fig 5). The DPPH radical scavenging activity of lysates was also lowered by treatment with oxidants alone but recovered when Cr was also present in the incubation medium (Fig 6).

Bottom Line: Recent reports have shown that Cr displays antioxidant activity which could explain its beneficial cellular effects.This was accompanied by decrease in glutathione levels.Creatinine, a by-product of Cr metabolism, was also shown to exert protective effects, although it was slightly less effective than Cr.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, Uttar Pradesh, India.

ABSTRACT
Creatine (Cr) is naturally produced in the body and stored in muscles where it is involved in energy generation. It is widely used, especially by athletes, as a staple supplement for improving physical performance. Recent reports have shown that Cr displays antioxidant activity which could explain its beneficial cellular effects. We have evaluated the ability of Cr to protect human erythrocytes and lymphocytes against oxidative damage. Erythrocytes were challenged with model oxidants, 2, 2'-azobis(2-amidinopropane) dihydrochloride (AAPH) and hydrogen peroxide (H2O2) in the presence and absence of Cr. Incubation of erythrocytes with oxidant alone increased hemolysis, methemoglobin levels, lipid peroxidation and protein carbonyl content. This was accompanied by decrease in glutathione levels. Antioxidant enzymes and antioxidant power of the cell were compromised while the activity of membrane bound enzyme was lowered. This suggests induction of oxidative stress in erythrocytes by AAPH and H2O2. However, Cr protected the erythrocytes by ameliorating the AAPH and H2O2 induced changes in these parameters. This protective effect was confirmed by electron microscopic analysis which showed that oxidant-induced cell damage was attenuated by Cr. No cellular alterations were induced by Cr alone even at 20 mM, the highest concentration used. Creatinine, a by-product of Cr metabolism, was also shown to exert protective effects, although it was slightly less effective than Cr. Human lymphocytes were similarly treated with H2O2 in absence and presence of different concentrations of Cr. Lymphocytes incubated with oxidant alone had alterations in various biochemical and antioxidant parameters including decrease in cell viability and induction of DNA damage. The presence of Cr attenuated all these H2O2-induced changes in lymphocytes. Thus, Cr can function as a blood antioxidant, protecting cells from oxidative damage, genotoxicity and can potentially increase their lifespan.

No MeSH data available.


Related in: MedlinePlus