title: Vitamin E protection of cell morphology under oxidative stress is related to cytoskeletal proteins in rat hepatocytes.
abstract: A significant change in cell morphology was observed in hepatocytes treated with t-butyl hydroperoxide (t-BH). This morphological change of multiple bleb formation on cell plasma membranes was related to cell damage, and the subsequent rupture of these blebs resulted in cell death. In cells incubated with alpha-tocopherol before t-BH treatment, bleb formation was significantly inhibited. Using fluorescence microscopy, actin organization was shown to be related to alpha-tocopherol status as demonstrated by early changes in the actin network of cells in the absence of alpha-tocopherol. Results from SDS-polyacrylamide gel electrophoresis further indicated that, under oxidative stress, actin molecules (45 kDa) decreased in amount and were accompanied by the formation of high molecular weight molecules. In the presence of the thiol reducing agent, dithiothreitol, both the decrease in monomeric actin and formation of high molecular weight molecules disappeared. The loss of actin showed a time-dependent response and could be observed after 15 min with t-BH treatment either in the presence or absence of alpha-tocopherol; however the extent was much more significant in cells with no alpha-tocopherol. Depletion of total membrane protein thiols was also related to vitamin E and was greater in cells with no alpha-tocopherol. The amount of cell damage, as determined by lactate dehydrogenase (LDH) leakage in cells with t-BH treatment over 120 min was decreased in the presence of alpha-tocopherol compared with the rapid increase of LDH leakage in the absence of alpha-tocopherol. These results indicate that vitamin E protection of cell morphology under oxidative stress is related to actin, with thiol groups in actin probably playing a key role.
authors.affiliation: Department of Nutrition, Chung Shan Medical College, Taichung, Taiwan, Republic of China.
authors.forename: J H
authors.initials: JH
authors.lastname: Kuo
authors.forename: H W
authors.initials: HW
authors.lastname: Chen
authors.forename: R G
authors.initials: RG
authors.lastname: Chou
authors.forename: C K
authors.initials: CK
authors.lastname: Lii
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002478:Cells, Cultured; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008856:Microscopy, Fluorescence; D018384:Oxidative Stress / Q000502:physiology*; D010545:Peroxides; D051381:Rats; D017207:Rats, Sprague-Dawley; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014810:Vitamin E / Q000494:pharmacology*; D020122:tert-Butylhydroperoxide
chemical_list: D000199:Actins; D000975:Antioxidants; D010545:Peroxides; D013438:Sulfhydryl Compounds; D014810:Vitamin E; D020122:tert-Butylhydroperoxide