title: Assessment of mitochondrial electron transport chain function in a primary astrocyte cell model of hyperhomocystinaemia.
abstract: Elevated plasma homocysteine (Hcy) has been detected in patients with various neurodegenerative conditions. Studies on neurones and cerebral tissue have revealed that hyperhomocystinaemia may inhibit mitochondrial electron transport chain (ETC) enzyme activity resulting in neuronal morbidity. As astrocytes convey a protective and supportive role towards neurones, we postulated that Hcy-induced astrocytic ETC inhibition may contribute to neurological dysfunction. In order to investigate this hypothesis, we established a cellular model of hyperhomocystinaemia using primary rat astrocytes. Which were incubated were incubated with 200 µM, 500 µM Hcy and the Hcy metabolite, thiolactone (10 µM). Following 96 h of incubation with 200 µM and 500 µM Hcy, an approximate two-fold (1.11 nmol/mg) and three-fold (1.45 nmol/mg) increase in mitochondrial levels of Hcy, respectively, were detected compared to control levels (0.54 nmol/mg). However, on exposure to Hcy (200 or 500 µM) and Hcy-thiolactone (10 µM), the activities of astrocytic ETC complex I, II-III and IV were found to be comparable to control levels. In addition, the extracellular lactate:pyruvate ratio and the intracellular glutathione status of primary rat astrocytes were not significantly different between Hcy (200 or 500 µM) treated and controls. In conclusion, the results of this study suggest that Hcy induced impairment of astrocytic ETC function may not contribute to the pathophysiology of hyperhomocystinaemia.
authors: Imperial College Medical School, London, UK.
authors: Fiona
authors: F
authors: Turkes
authors: Elaine
authors: E
authors: Murphy
authors: John
authors: J
authors: Land
authors: Berna
authors: B
authors: Demiray
authors: Kate
authors: K
authors: Duberley
authors: Antony
authors: A
authors: Briddon
authors: Iain
authors: I
authors: Hargreaves
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D045222:Electron Transport Chain Complex Proteins / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D006710:Homocysteine / Q000378:metabolism / Q000633:toxicity*; D020138:Hyperhomocysteinemia / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D008954:Models, Biological; D061251:Primary Cell Culture; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
chemical_list: D045222:Electron Transport Chain Complex Proteins; D006710:Homocysteine; D005978:Glutathione