PURPOSE: To use different paradigms of oxidative and metabolic stress in a cellular model of Leber hereditary optic neuropathy (LHON), with the aim of evaluating the efficacy of potentially therapeutic molecules for the treatment of this disease. METHODS: Cybrids bearing one of the three most common LHON pathogenic mutations (11778/ND4, 3460/ND1, 14484/ND6) were incubated with two compounds known to induce oxidative injury, tert-butyl hydroperoxide (t-BH) and rotenone. To mimic metabolic stress, cells were incubated in a glucose-free medium containing galactose. Cell viability was determined using the MTT assay. To identify the apoptotic type of cell death, nuclear morphology was examined after cell loading with Hoechst. Cellular glutathione (GSH), and oxidized glutathione (GSSG) levels were measured enzymatically. RESULTS: Incubation with t-BH caused apoptotic cell death of control and LHON cybrids, whereas only LHON cybrids were damaged by rotenone concentrations up to 2.5 muM. Both types of stress caused a marked imbalance in the glutathione levels, but an increase in the GSSG/GSH+GSSG ratio was detected only after rotenone treatment. The efficacy of several antioxidant and antiapoptotic compounds was then assessed in cells exposed to these two oxidative paradigms. Only exogenous GSH remarkably protected the t-BH- and rotenone-treated cybrids from cell death. In contrast, GSH was unable to increase the viability of cybrids exposed to metabolic stress. CONCLUSIONS: These results suggest that GSH is an effective antioxidant compound to be tested as a potential treatment for LHON.
PURPOSE: To use different paradigms of oxidative and metabolic stress in a cellular model of Leber hereditary optic neuropathy (LHON), with the aim of evaluating the efficacy of potentially therapeutic molecules for the treatment of this disease. METHODS: Cybrids bearing one of the three most common LHON pathogenic mutations (11778/ND4, 3460/ND1, 14484/ND6) were incubated with two compounds known to induce oxidative injury, tert-butyl hydroperoxide (t-BH) and rotenone. To mimic metabolic stress, cells were incubated in a glucose-free medium containing galactose. Cell viability was determined using the MTT assay. To identify the apoptotic type of cell death, nuclear morphology was examined after cell loading with Hoechst. Cellular glutathione (GSH), and oxidized glutathione (GSSG) levels were measured enzymatically. RESULTS: Incubation with t-BH caused apoptotic cell death of control and LHON cybrids, whereas only LHON cybrids were damaged by rotenone concentrations up to 2.5 muM. Both types of stress caused a marked imbalance in the glutathione levels, but an increase in the GSSG/GSH+GSSG ratio was detected only after rotenone treatment. The efficacy of several antioxidant and antiapoptotic compounds was then assessed in cells exposed to these two oxidative paradigms. Only exogenous GSH remarkably protected the t-BH- and rotenone-treated cybrids from cell death. In contrast, GSH was unable to increase the viability of cybrids exposed to metabolic stress. CONCLUSIONS: These results suggest that GSH is an effective antioxidant compound to be tested as a potential treatment for LHON.
Authors: Anna Ghelli; Concetta V Tropeano; Maria Antonietta Calvaruso; Alessandra Marchesini; Luisa Iommarini; Anna Maria Porcelli; Claudia Zanna; Vera De Nardo; Andrea Martinuzzi; Flemming Wibrand; John Vissing; Ivana Kurelac; Giuseppe Gasparre; Nur Selamoglu; Fevzi Daldal; Michela Rugolo Journal: Hum Mol Genet Date: 2013-02-14 Impact factor: 6.150
Authors: Chun Shi Lin; Mark S Sharpley; Weiwei Fan; Katrina G Waymire; Alfredo A Sadun; Valerio Carelli; Fred N Ross-Cisneros; Peter Baciu; Eric Sung; Meagan J McManus; Billy X Pan; Daniel W Gil; Grant R Macgregor; Douglas C Wallace Journal: Proc Natl Acad Sci U S A Date: 2012-11-05 Impact factor: 11.205
Authors: Anna Ghelli; Anna Maria Porcelli; Claudia Zanna; Sara Vidoni; Stefano Mattioli; Anna Barbieri; Luisa Iommarini; Maria Pala; Alessandro Achilli; Antonio Torroni; Michela Rugolo; Valerio Carelli Journal: PLoS One Date: 2009-11-19 Impact factor: 3.240