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脂质过氧化与生物源醛类:从4-羟基壬烯醛的鉴定到生物病理学的进一步进展

Lipid peroxidation and biogenic aldehydes: from the identification of 4-hydroxynonenal to further achievements in biopathology.

作者信息

Comporti M

机构信息

Instituto di Patologia Generale, Università di Siena, Italy.

出版信息

Free Radic Res. 1998 Jun;28(6):623-35. doi: 10.3109/10715769809065818.

Abstract

The formation, reactivity and toxicity of aldehydes originating from lipid peroxidation of cellular membranes are reviewed. Very reactive aldehydes, namely 4-hydroxyalkenals, were first shown to be formed in autoxidizing chemical systems. It was subsequently shown that 4-hydroxyalkenals are formed in biological conditions, i.e. during lipid peroxidation of liver microsomes incubated in the NADPH-Fe systems. Our studies carried out in collaboration with Hermann Esterbauer which led to the identification of 4-hydroxynonenal (4-HNE) are reported. 4-HNE was the most cytotoxic aldehyde and was then assumed as a model molecule of oxidative stress. Many other aldehydes (alkanals, alk-2-enals and dicarbonyl compounds) were then identified in peroxidizing liver microsomes or hepatocytes. The in vivo formation of aldehydes in liver of animals intoxicated with agents that promote lipid peroxidation was shown in further studies. In a first study, evidence was forwarded for aldehydes (very likely alkenals) bound to liver microsomal proteins of CCl4 or BrCCl3-intoxicated rats. In a second study, 4-HNE and a number of other aldehydes (alkanals and alkenals) were identified in the free (non-protein bound) form in liver extracts from bromobenzene or allyl alcohol-poisoned mice. The detection of free 4-HNE in the liver of CCl(4) or BrCC1(3)-poisoned animals was obtained with the use of an electrochemical detector, which greatly increased the sensitivity of the HPLC method. Furthermore, membrane phospholipids bearing carbonyl groups were demonstrated in both in vitro (incubation of microsomes with NADPH-Fe) and in vivo (CC1(4) or BrCCl(3) intoxication) conditions. Finally, the results concerned with the histochemical detection of lipid peroxidation are reported. The methods used were based on the detection of lipid peroxidation-derived carbonyls. Very good results were obtained with the use of fluorescent reagents for carbonyls, in particular with 3-hydroxy-2-naphtoic acid hydrazide (NAH) and analysis with confocal scanning fluorescence microscopy with image video analysis. The significance of formation of toxic aldehydes in biological membranes is discussed.

摘要

本文综述了细胞膜脂质过氧化产生的醛类的形成、反应活性和毒性。极具反应活性的醛类,即4-羟基烯醛,最初是在自氧化化学体系中被发现的。随后有研究表明,4-羟基烯醛在生物条件下也会形成,即在NADPH-Fe体系中孵育的肝微粒体脂质过氧化过程中。本文报道了我们与赫尔曼·埃斯特鲍尔合作开展的研究,该研究最终鉴定出了4-羟基壬烯醛(4-HNE)。4-HNE是细胞毒性最强的醛类,因此被视为氧化应激的模型分子。随后,人们在过氧化的肝微粒体或肝细胞中又鉴定出了许多其他醛类(烷醛、烯-2-醛和二羰基化合物)。进一步的研究表明,在用促进脂质过氧化的试剂处理的动物肝脏中,醛类会在体内形成。在第一项研究中,有证据表明,四氯化碳(CCl4)或三氯溴甲烷(BrCCl3)中毒大鼠的肝微粒体蛋白结合了醛类(很可能是烯醛)。在第二项研究中,在溴苯或烯丙醇中毒小鼠的肝脏提取物中,鉴定出了游离(非蛋白结合)形式的4-HNE和其他一些醛类(烷醛和烯醛)。使用电化学检测器实现了对CCl4或BrCCl3中毒动物肝脏中游离4-HNE的检测,这大大提高了高效液相色谱法(HPLC)的灵敏度。此外,在体外(微粒体与NADPH-Fe孵育)和体内(CCl4或BrCCl3中毒)条件下,均证实了带有羰基的膜磷脂的存在。最后,报道了与脂质过氧化组织化学检测相关的结果。所采用的方法基于对脂质过氧化衍生羰基的检测。使用羰基荧光试剂,特别是3-羟基-2-萘甲酸酰肼(NAH),并结合共聚焦扫描荧光显微镜和图像视频分析进行分析,取得了很好的结果。本文还讨论了生物膜中有毒醛类形成的意义。

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