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不同氧化敏感型酵母菌种中自发的和自由基诱导的质膜脂质过氧化作用及其抗氧化剂抑制作用

Spontaneous and radical-induced plasma membrane lipid peroxidation in differently oxidant-sensitive yeast species and its suppression by antioxidants.

作者信息

Krasowska A, Lukaszewicz M, Oświecimska M, Witek S, Sigler K

机构信息

Institute of Microbiology, University of Wrocław, 51 148 Wrocław, Poland.

出版信息

Folia Microbiol (Praha). 2000;45(6):509-14. doi: 10.1007/BF02818719.

Abstract

Formation of thiobarbituric acid-reactive substances (TBRS; nmol/mg lipids) indicative of lipid peroxidation was measured in whole cells and in isolated plasma membrane lipids from three yeast species differing in oxidant sensitivity (Schizosaccharomyces pombe, Saccharomyces cerevisiae and Rhodotorula glutinis) after exposure to the Fenton reagent, FeII, H2O2, tert-butyl hydroperoxide (TBHP) and azo compounds (AAPH, ACHN). In whole cells, spontaneous TBRS formation rose in the sequence S. pombe < S. cerevisiae < R. glutinis (1:approximately 5:approximately 7). Oxidants increased the TBRS production 13-18 fold in the sequence FeII approximately TBHP > AAPH approximately ACHN approximately Fe-Fenton > H2O2. This increase need not be solely due to increased lipid peroxidation. In isolated plasma membrane lipids from all three species, the spontaneous TBRS production referred to 1 mg lipids was 9-13-fold higher than in whole cells. In S. pombe lipids, only TBHP increased the TBRS production. In lipids from S. cerevisiae and R. glutinis, all added oxidants increased the spontaneous TBRS production 2-3 times in the sequence TBHP > ACHN > AAPH > FeII > Fe-Fenton > H2O2. Oxidant-induced TBRS production in both whole cells and isolated membrane lipids was partially suppressed by the lipid peroxidation inhibitors 2,6-di-tert-butyl-4-methylphenol ("butylated hydroxytoluene"; BHT) and the newly synthesized PYA12 compound. Both agents were more effective in isolated lipids than in whole cells and against OH.-producing than against ROO.- or RO.-producing oxidants. Yeast membrane lipids, which are generally poor in polyunsaturated fatty acids, are thus subject to perceptible lipid peroxidation.

摘要

在暴露于芬顿试剂、亚铁离子(FeII)、过氧化氢(H2O2)、叔丁基过氧化氢(TBHP)和偶氮化合物(AAPH、ACHN)后,对三种氧化敏感性不同的酵母(粟酒裂殖酵母、酿酒酵母和粘红酵母)的全细胞以及分离的质膜脂质中硫代巴比妥酸反应性物质(TBRS;纳摩尔/毫克脂质)的形成进行了测定,该物质可指示脂质过氧化。在全细胞中,自发TBRS形成按粟酒裂殖酵母<酿酒酵母<粘红酵母的顺序增加(1:约5:约7)。氧化剂使TBRS产量按亚铁离子≈叔丁基过氧化氢>偶氮二异丁腈≈偶氮环己烷甲腈>铁-芬顿>过氧化氢的顺序增加13 - 18倍。这种增加不一定完全是由于脂质过氧化增加所致。在所有三种酵母的分离质膜脂质中,以1毫克脂质计的自发TBRS产量比全细胞中的高9 - 13倍。在粟酒裂殖酵母脂质中,只有叔丁基过氧化氢增加了TBRS产量。在酿酒酵母和粘红酵母的脂质中,所有添加的氧化剂使自发TBRS产量按叔丁基过氧化氢>偶氮环己烷甲腈>偶氮二异丁腈>亚铁离子>铁-芬顿>过氧化氢的顺序增加2 - 3倍。脂质过氧化抑制剂2,6 - 二叔丁基-4 - 甲基苯酚(“丁基化羟基甲苯”;BHT)和新合成的PYA12化合物部分抑制了全细胞和分离膜脂质中氧化剂诱导的TBRS产生。这两种试剂在分离脂质中比在全细胞中更有效,并且对产生羟基自由基的氧化剂比对产生过氧自由基或烷氧自由基的氧化剂更有效。酵母膜脂质通常富含多不饱和脂肪酸,因此会发生明显的脂质过氧化。

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