The use of xenobiotic-mediated methaemoglobin formation to assess the effects of thyroid hormones on diabetic and non-diabetic human erythrocytic oxidant defence mechanisms in vitro

Diabetes is associated with an abnormal incidence of hypothyroidism, which exacerbates hyperglycaemia, so further damaging already compromised erythrocytic defence mechanisms. Methaemoglobin formation is a useful measure of the health of these mechanisms, as it determines the resistance of diabetic erythrocytes to sustained oxidative stress. The effect of L-tri-iodothyronine (T ₃) was, therefore, studied on nitrite and monoacetyldapsone hydroxylamine (MADDS-NHOH) mediated methaemoglobin formation in diabetic and non-diabetic human erythrocytes. Diabetic erythrocytes showed less sensitivity compared with non-diabetics to methaemoglobin formation mediated by both compounds. A 30 min pre-incubation with T ₃ at 3 and 30 nM did not affect nitrite-mediated methaemoglobin formation compared with control observations in both cell types. In diabetic erythrocytes incubated with T ₃ at 30 nM, there were significant increases in MADDS-NHOH-mediated methaemoglobin formation compared with control in both diabetic and non-diabetic cells. Studies comparing blood isolated from diabetic patients stabilised on thyroxine (T ₄; 50 μG/day), T ₄-free diabetics and non-diabetics, showed that T ₄ supplementation significantly increased MADDS-NHOH-mediated methaemoglobin formation compared with T ₄-free diabetic cells so that for two time points, T ₄-treated diabetic erythrocytic methaemoglobin formation was indistinguishable from that of non-diabetics. These studies indicate that T ₄ supplementation improves some erythrocytic oxidant defence mechanisms in a time dependent manner. © 2002 Elsevier Science B.V. All rights reserved.