The Antioxidant and Pro-oxidant Impacts of Varying Levels of Alpha-Lipoic Acid on Biomarkers of Myoglobin Oxidation in Vitro

Amani Ibrahim  Farah; Mousa Numan  Ahmad; Tareq Musbah  Al-Qirim

doi:10.35516/jjas.v16i4.63

Authors

Amani Ibrahim Farah University of Jordan, Amman, Jordan
Mousa Numan Ahmad University of Jordan, Amman, Jordan
Tareq Musbah Al-Qirim Amman , Jordan

DOI:

https://doi.org/10.35516/jjas.v16i4.63

Keywords:

Antioxidants, pro-oxidant, Lipoic acid, Myoglobin, Protein carbonyls, Protein-free thiols, Chronic diseases

Abstract

R-alpha-lipoic acid (R-ALA) has been known to protect protein oxidation and lessen the pathogenesis of oxidative-related multiple diseases; however, its dosing remains unresolved. This study aimed to examine whether in vitro R-ALA varying levels would have antioxidant or pro-oxidant impacts on biomarkers of myoglobin oxidation in terms of carbonyls and free thiols for myoglobin upon long-term incubation. Myoglobin (1mg/mL) was concentrated with 6 different concentrations of R-ALA: 50 µM, 100 µM, 500 µM, 1mM, 2mM and 4mM for 30 days at pH 6.6 and temperature 37 °C. Myoglobin oxidative modifications as protein carbonyls and its oxidative defense as free thiols were determined by standard procedures. Thirty-day coincubation of native myoglobin with R-ALA at 500 µM, 1mM, 2mM, and 4mM significantly (p<0.05) elevated carbonyls (2.51±0.19; 2.59±0.22; 2.71±0.32 and 2.79±0.39 nmol/ mg protein respectively) compared to their levels in native control myoglobin (1.67±0.43 nmol/ mg protein) and significantly (p<0.05) decreased free thiols (4.60±0.36; 4.49±0.46; 4.38±0.28 and 4.07±0.39 nmol/ mg protein respectively) against their levels in native control myoglobin (5.71±0.62 nmol/ mg protein). Conversely, coincubation of myoglobin with 50µM and 100µM R-ALA reduced carbonyls (1.02±0.29 and 0.9±0.19 nmol/ mg protein respectively) compared to the control levels (1.67±0.43 nmol/ mg protein) and elevated free thiols (6.1±0.28 and 6.83±0.28 nmol/ mg protein respectively) against control levels (5.71±0.62 nmol/ mg protein) levels; 100µM elicited significant (p<0.05) differences, but 50µM did not. Findings indicate that high levels of R-ALA (0.5-4mM) provoked myoglobin oxidative damage while moderate levels (50-100µM) protected protein upon any spontaneous oxidative damage during long-term coincubation. Thus, R-ALA concentrations, which set the balance between R-ALA pro- and antioxidants, dictate the primary impacts of R-ALA on myoglobin redox status. Additional in vivo investigations are needed to assess the therapeutic insights of current findings.

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Author Biographies

Amani Ibrahim Farah, University of Jordan, Amman, Jordan

Department of Nutrition and Food Technology, Human Nutrition and Dietetics, University of Jordan, Amman 11942, Jordan

Mousa Numan Ahmad, University of Jordan, Amman, Jordan

Department of Nutrition and Food Technology, Human Nutrition and Dietetics

Tareq Musbah Al-Qirim, Amman , Jordan

Faculty of Pharmacy, Al-Zaytoonah University of Jordan

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