title: Study the oxidative injury of yeast cells by NADH autofluorescence.
abstract: Autofluorescence has an advantage over the extrinsic fluorescence of an unperturbed environment during investigation, especially in complex system such as biological cells and tissues. NADH is an important fluorescent substance in living cells. The time courses of intracellular NADH autofluorescence in the process of yeast cells exposed to H(2)O(2) and ONOO(-) have been recorded in detail in this work. In the presence of different amounts of H(2)O(2) and ONOO(-), necrosis, apoptosis and reversible injury are initiated in yeast cells, which are confirmed by acridine orange/ethidum bromide and Annexin V/propidium iodide staining. It is found that intracellular NADH content increases momently in the beginning of the apoptotic process and then decreases continually till the cell dies. The most remarkable difference between the apoptotic and the necrotic process is that the NADH content in the latter case changes much more sharply. Further in the case of reversible injury, the time course of intracellular NADH content is completely different from the above two pathways of cell death. It just decreases to some degree firstly and then resumes to the original level. Based on the role of NADH in mitochondrial respiratory chain, the time course of intracellular NADH content is believed to have reflected the response of mitochondrial redox state to oxidative stress. Thus, it is found that the mitochondrial redox state changes differently in different pathways of oxidative injury in yeast cells.
authors.affiliation: Department of Chemistry, Wuhan University, Wuhan 430072, China.
authors.forename: Ju
authors.initials: J
authors.lastname: Liang
authors.forename: Wen-Lan
authors.initials: WL
authors.lastname: Wu
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Liu
authors.forename: Yun-Jun
authors.initials: YJ
authors.lastname: Mei
authors.forename: Ru-Xiu
authors.initials: RX
authors.lastname: Cai
authors.forename: Ping
authors.initials: P
authors.lastname: Shen
mesh_terms: D017209:Apoptosis; D005453:Fluorescence; D006861:Hydrogen Peroxide / Q000633:toxicity; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D009243:NAD / Q000737:chemistry*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000633:toxicity; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013481:Superoxides / Q000633:toxicity
chemical_list: D017382:Reactive Oxygen Species; D009243:NAD; D013481:Superoxides; D006861:Hydrogen Peroxide