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. 2012;10(1):1-10.
doi: 10.2203/dose-response.10-002.Siddique. Epub 2011 Aug 11.

Estimation of lipid peroxidation induced by hydrogen peroxide in cultured human lymphocytes

Yasir Hasan Siddique  1 , Gulshan Ara, Mohammad Afzal
Affiliations

Estimation of lipid peroxidation induced by hydrogen peroxide in cultured human lymphocytes

Yasir Hasan Siddique et al. Dose Response. 2012.

Abstract

Malondialdehyde (MDA) is used for the estimation of damage by reactive oxygen species. MDA is a major reactive aldehyde resulting from the peroxidation of biological membranes. The most common method used to assess MDA production is the thiobarbituric acid (TBARS) assay. However, the value of this method is curbed by low specificity and has been criticized for its use in human studies. In the present study we have used an alternative method for the estimation of MDA production i.e. reaction of MDA with a chromogenic agent 1-methyl-2-phenylindole at 45°C. The paper describes the method of preparing standards for the estimation of MDA (lipid peroxidation) after the treatment with an oxidative stress inducing agent hydrogen peroxide (H(2)O(2)). In the present study, the treatments of 1, 5, 10, 20, 50, 100, 150 and 200 μM of H(2)O(2) induced significant increase in lipid peroxidation as compared to the untreated ones. The results suggest that the present method can be used to measure the lipid peroxidation in cultured human peripheral blood lymphocytes and is specific for MDA estimation.

Keywords: human lymphocytes; hydrogen-peroxide; lipid peroxidation; malonaldehyde; oxidative stress.

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Figures

FIG. 1.
FIG. 1.
Steps of lipid peroxidation (Alessio, 2000).
FIG. 2.
FIG. 2.
Standard graph for the estimation of lipid peroxidation in cultured human peripheral blood lymphocytes.
FIG. 3.
FIG. 3.
Lipid peroxidation after 24 h treatment of the H2O2 on cultured human lymphocytes (*p<0.05).
FIG. 4.
FIG. 4.
Lipid peroxidation after 48h treatment of the H2O2 on cultured human lymphocytes (*p<0.05).
FIG. 5.
FIG. 5.
Effect of hydrogen peroxide treatment on the mean absorbance at 586 nm in cultured human peripheral blood lymphocytes after 24 h.
FIG. 6.
FIG. 6.
Effect of hydrogen peroxide treatment on the mean absorbance at 586 nm in cultured human peripheral blood lymphocytes after 48 h.
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