Synthesis and Cellular Effects of an Intracellularly Activated Analogue of 4-Hydroxynonenal

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• 作者：M. Diana Neely ; Venkataraman Amarnath ; Carl Weitlauf ; and Thomas J. Montine
• 刊名：Chemical Research in Toxicology
• 出版年：2002
• 出版时间：January 2002
• 年：2002
• 卷：15
• 期：1
• 页码：40 - 47
• 全文大小：166K
• 年卷期：v.15,no.1(January 2002)
• ISSN：1520-5010

文摘

4-Hydroxy-2-nonenal (HNE) has been recognized as reactive product of lipid peroxidationand has been suggested to play a role in the pathogenesis in several common diseases as wellas injuries caused by environmental toxicants. Although formed intracellularly in vivo, forpractical reasons this molecule is applied extracellularly in order to analyze its biological effects.The focus of this study was to develop an approach that would enable intracellular HNEproduction in the absence of the many other products and processes that occur in cellsexperiencing generalized oxidative stress. To this end, we synthesized 1,1,4-tris(acetyloxy)-2(E)-nonene (HNE[Ac]₃), a triester analogue of HNE that is itself unreactive but could behydrolyzed intracellularly presumably by lipases and/or esterases into the highly reactive HNE.In vitro lipase rapidly converted HNE(Ac)₃ initially to 4-acetyloxy-2-nonenal (HNE[Ac]₁) andthen to HNE. Neuro 2A cell lysate also caused a rapid hydrolysis of HNE(Ac)₃ into HNE(Ac)₁and HNE. Incubation of BSA with HNE(Ac)₃ resulted in protein-adduct formation only inthe presence of lipase. We demonstrated adduction of HNE to proteins in Neuro 2A cells exposedto HNE(Ac)₃ by immunoblotting and immunocytochemistry using antibodies specific for HNE-Michael adducts on proteins. We have previously shown that microtubule organization is verysensitive to HNE. Analysis of Neuro 2A cell microtubules showed that this cytoplasmic organelleis similarly sensitive to HNE and HNE(Ac)₃.