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An iron-based beverage, HydroFerrate fluid (MRN-100), alleviates oxidative stress in murine lymphocytes in vitro.

Ghoneum M, Matsuura M, Gollapudi S - Nutr J (2009)

Bottom Line: Results show, as expected, that culture of splenic cells with H(2)O(2) alone results in a significant increase in cell death (apoptosis) as compared to control (CM) cells.In contrast, pre-treatment of cells with MRN-100 followed by H(2)O(2) treatment results in significantly reduced levels of apoptosis.In addition, MRN-100 partially prevents H(2)O(2) -induced down-regulation of the anti-apoptotic molecule Bcl-2 and upregulation of the pro-apoptotic molecule Bax.Our findings suggest that MRN-100 may offer a protective effect against oxidative stress-induced apoptosis in lymphocytes.

View Article: PubMed Central - HTML - PubMed

Affiliation: Charles Drew University of Medicine and Science, Department of Otolaryngology, Los Angeles, California 90059, USA. mghoneum@ucla.edu

ABSTRACT

Background: Several studies have examined the correlation between iron oxidation and H(2)O(2) degradation. The present study was carried out to examine the protective effects of MRN-100 against stress-induced apoptosis in murine splenic cells in vitro. MRN-100, or HydroFerrate fluid, is an iron-based beverage composed of bivalent and trivalent ferrates.

Methods: Splenic lymphocytes from mice were cultured in the presence or absence of MRN-100 for 2 hrs and were subsequently exposed to hydrogen peroxide (H(2)O(2) ) at a concentration of 25 microM for 14 hrs. Percent cell death was examined by flow cytometry and trypan blue exclusion. The effect of MRN-100 on Bcl-2 and Bax protein levels was determined by Western blot.

Results: Results show, as expected, that culture of splenic cells with H(2)O(2) alone results in a significant increase in cell death (apoptosis) as compared to control (CM) cells. In contrast, pre-treatment of cells with MRN-100 followed by H(2)O(2) treatment results in significantly reduced levels of apoptosis.In addition, MRN-100 partially prevents H(2)O(2) -induced down-regulation of the anti-apoptotic molecule Bcl-2 and upregulation of the pro-apoptotic molecule Bax.

Conclusion: Our findings suggest that MRN-100 may offer a protective effect against oxidative stress-induced apoptosis in lymphocytes.

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Cell death analysis of splenic lymphocytes by trypan blue assay. Splenic lymphocytes were cultured in the presence of H2O2, MRN-100, MRN-100 + H2O2, or control (CM). At 16 hrs, cells from the different groups were treated with trypan blue stain for 5 minutes and counted. The percent of dead cells was calculated out of a total of 300 cells. * indicates p < 0.02 as compared to cells treated with H2O2. Data shown is representative of three independent experiments.
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Figure 2: Cell death analysis of splenic lymphocytes by trypan blue assay. Splenic lymphocytes were cultured in the presence of H2O2, MRN-100, MRN-100 + H2O2, or control (CM). At 16 hrs, cells from the different groups were treated with trypan blue stain for 5 minutes and counted. The percent of dead cells was calculated out of a total of 300 cells. * indicates p < 0.02 as compared to cells treated with H2O2. Data shown is representative of three independent experiments.

Mentions: The protective effect of MRN-100 against H2O2-induced cell death was further confirmed by trypan blue exclusion tests. Results in Figure 2 show a high level of dead splenic cells post-treatment with H2O2. In contrast, pre-treatment of splenic cells with MRN-100 prevents H2O2-induced cell death.


An iron-based beverage, HydroFerrate fluid (MRN-100), alleviates oxidative stress in murine lymphocytes in vitro.

Ghoneum M, Matsuura M, Gollapudi S - Nutr J (2009)

Cell death analysis of splenic lymphocytes by trypan blue assay. Splenic lymphocytes were cultured in the presence of H2O2, MRN-100, MRN-100 + H2O2, or control (CM). At 16 hrs, cells from the different groups were treated with trypan blue stain for 5 minutes and counted. The percent of dead cells was calculated out of a total of 300 cells. * indicates p < 0.02 as compared to cells treated with H2O2. Data shown is representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2683876&req=5

Figure 2: Cell death analysis of splenic lymphocytes by trypan blue assay. Splenic lymphocytes were cultured in the presence of H2O2, MRN-100, MRN-100 + H2O2, or control (CM). At 16 hrs, cells from the different groups were treated with trypan blue stain for 5 minutes and counted. The percent of dead cells was calculated out of a total of 300 cells. * indicates p < 0.02 as compared to cells treated with H2O2. Data shown is representative of three independent experiments.
Mentions: The protective effect of MRN-100 against H2O2-induced cell death was further confirmed by trypan blue exclusion tests. Results in Figure 2 show a high level of dead splenic cells post-treatment with H2O2. In contrast, pre-treatment of splenic cells with MRN-100 prevents H2O2-induced cell death.

Bottom Line: Results show, as expected, that culture of splenic cells with H(2)O(2) alone results in a significant increase in cell death (apoptosis) as compared to control (CM) cells.In contrast, pre-treatment of cells with MRN-100 followed by H(2)O(2) treatment results in significantly reduced levels of apoptosis.In addition, MRN-100 partially prevents H(2)O(2) -induced down-regulation of the anti-apoptotic molecule Bcl-2 and upregulation of the pro-apoptotic molecule Bax.Our findings suggest that MRN-100 may offer a protective effect against oxidative stress-induced apoptosis in lymphocytes.

View Article: PubMed Central - HTML - PubMed

Affiliation: Charles Drew University of Medicine and Science, Department of Otolaryngology, Los Angeles, California 90059, USA. mghoneum@ucla.edu

ABSTRACT

Background: Several studies have examined the correlation between iron oxidation and H(2)O(2) degradation. The present study was carried out to examine the protective effects of MRN-100 against stress-induced apoptosis in murine splenic cells in vitro. MRN-100, or HydroFerrate fluid, is an iron-based beverage composed of bivalent and trivalent ferrates.

Methods: Splenic lymphocytes from mice were cultured in the presence or absence of MRN-100 for 2 hrs and were subsequently exposed to hydrogen peroxide (H(2)O(2) ) at a concentration of 25 microM for 14 hrs. Percent cell death was examined by flow cytometry and trypan blue exclusion. The effect of MRN-100 on Bcl-2 and Bax protein levels was determined by Western blot.

Results: Results show, as expected, that culture of splenic cells with H(2)O(2) alone results in a significant increase in cell death (apoptosis) as compared to control (CM) cells. In contrast, pre-treatment of cells with MRN-100 followed by H(2)O(2) treatment results in significantly reduced levels of apoptosis.In addition, MRN-100 partially prevents H(2)O(2) -induced down-regulation of the anti-apoptotic molecule Bcl-2 and upregulation of the pro-apoptotic molecule Bax.

Conclusion: Our findings suggest that MRN-100 may offer a protective effect against oxidative stress-induced apoptosis in lymphocytes.

Show MeSH
Related in: MedlinePlus