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. 2013;17(1):22-8.
doi: 10.6091/ibj.1092.2012.

Antioxidant effects of vitamins C and e on the low-density lipoprotein oxidation mediated by myeloperoxidase

Seyed Ziyae Aldin Samsam Shariat  1 , Sayed Abolfazl Mostafavi  1 , Farzad Khakpour  1
Affiliations

Antioxidant effects of vitamins C and e on the low-density lipoprotein oxidation mediated by myeloperoxidase

Seyed Ziyae Aldin Samsam Shariat et al. Iran Biomed J. 2013.

Abstract

Background: Oxidative modification of low-density lipoprotein (LDL) appears to be an early step in the pathogenesis of atherosclerosis. Meanwhile, myeloperoxidase (MPO)-catalyzed reaction is one of the potent pathway for LDL oxidation in vivo. The aim of this study was to evaluate in vitro antioxidant effects of vitamins C and E on LDL oxidation mediated by MPO.

Methods: MPO was isolated from fresh plasma by sequential centrifugation using density ultracentrifugation. It was incubated with LDL and the LDL oxidation level was determined spectrophotometrically by measuring conjugated diene absorbance at 234 nm. Furthermore, vitamin C (50-200 mM) and vitamin E (10-40 mM) were added and the LDL oxidation level was determined.

Results: The purity index of MPO and its enzymatic activity were 0.69 and 1127 U/mg protein, respectively. It was demonstrated that vitamin C in vitro inhibited LDL oxidation mediated by MPO; however, vitamin E was unable to act in the same way. The protection by vitamin C was concentration dependent and maximum protective effect of vitamin C was observed at 150 mM, where about 64% of the LDL oxidation was inhibited. Vitamin C increased lag time of LDL oxidation mediated by MPO up to 2.4 times.

Conclusion: It can be concluded from our results that vitamin C is able to improve LDL resistance to oxidative modification in vitro. In addition, vitamin C might be effective in LDL oxidation mediated by MPO in vivo, resulting in reduction of atherosclerosis process rate.

Keywords: Antioxidant; Myeloperoxidase; Low-density lipoprotein; Vitamin E.

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Figures

Fig. 1
Fig. 1
Effect of different concentrations of vitamin C on LDL oxidation induced by myeloperoxidase (MPO) system. Low-density lipoprotein (LDL) was incubated in the absence and presence of 50-200 mM of vitamin C and changes in absorbance at 234 nm were read at intervals of 5 min for a period of 400 min. values represents mean   SD of three independent experiments. vit, vitamin
Fig. 2
Fig. 2
Effects of different concentrations of vitamin C on lag time of low-density lipoprotein (LDL) oxidation induced by myeloperoxidase system. Values were obtained from LDL oxidation curves. Values represent mean   SD of three independent experiments
Fig. 3
Fig. 3
Effect of different concentrations of vitamin C on conjugated dienes induced by myeloperoxidase-system. Conjugated dienes produced were calculated from the maximum increase of the absorbance and molar absorbance for conjugated dienes of 2.95 × 104M-1cm-1.
Fig. 4
Fig. 4
Effect of different concentrations of vitamin E ( -tocopherol) on low-density lipoprotein (LDL) oxidation induced by myeloperoxidase (MPO)/H2O2/Cl- system. LDL was incubated in the absence and presence of 10-40 mM of vitamin E, and changes in absorbance at 234 nm were read at intervals of 5 min for a period of 400 min. Values represents mean   SD of three independent experiments. Vit, vitamin
Fig. 5
Fig. 5
Effect of different concentrations of vitamin E on lag time of the low-density lipoprotein (LDL) oxidation mediated by myeloperoxidase. Values were obtained from the LDL oxidation curves of Figure 4. Values represent mean   SD of three independent experiments.
Fig. 6
Fig. 6
Effect of different concentrations of vitamin E on the conjugated dienes produced during low-density lipoprotein (LDL) oxidation mediated by myeloperoxidase. Values were obtained from the LDL oxidation curves of Figure 4. Values represent mean   SD of three independent experiments.
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