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Monitoring antioxidant effects using methaemoglobin formation in diabetic erythrocytes

School of Pharmacy, Aston University, Birmingham, UK, m.d.coleman{at}aston.ac.uk

Helen L Tolley

School of Pharmacy, Aston University, Birmingham, UK

Amisha K Desai

School of Pharmacy, Aston University, Birmingham, UK

Deficiencies in the reducing power of diabetic erythrocytes can be studied using the sulphone metabolite, monoacetyl dapsone hydroxylamine (MADDS-NHOH) to generate methaemoglobin, which is a form of oxidative stress. The effects of three antioxidants, dyhydrolipoic acid (DHLA), lipoic acid and ascorbate were compared using this method in erythrocytes of type 1 diabetic patients. During 2-hour incubations, DHLA and lipoic acid significantly reduced MADDS-NHOH-mediated methaemoglobin formation to the same extent in diabetic and non-diabetic erythrocytes. In contrast, ascorbate markedly increased MADDS-NHOH-mediated methaemoglobin formation at all time points up to 2 hours for diabetic cells and 15—60 minutes in non-diabetic erythrocytes. Ascorbate abolished the deficiency in methaemoglobin sensitivity normally seen in diabetic erythrocytes compared with non-diabetic erythrocytes. During a 1-hour pre-incubation period, neither DHLA, lipoic acid nor ascorbate significantly altered total thiol concentrations. After a 2-hour incubation with MADDS-NHOH, thiol concentrations did not significantly change in the non-diabetic erythrocytes, but they fell significantly in the diabetic cells. Diabetic and non-diabetic erythrocytes pre-incubated with ascorbate, DHLA and lipoic acid showed no changes in thiol levels in the presence of MADDS-NHOH. In summary, ascorbate, DHLA and lipoic acid showed contrasting effects on methaemoglobin generation, although they each abolished the diabetic erythrocytic deficit in total thiol status caused by hydroxylamine-mediated methaemoglobin formation. This work provides evidence for the potential future use of antioxidant supplements in diabetic management.

Key Words: diabetes • methaemoglobin • antioxidant • erythrocyte • human.

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The British Journal of Diabetes & Vascular Disease, Vol. 1, No. 1, 88-92 (2001)
DOI: 10.1177/14746514010010011601


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This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Coleman, M. D Articles by Desai, A. K Search for Related Content Social Bookmarking                         What's this?