HOCl-induced lysophospholipid generation from diacyl- and alkenyl-acyl-phospholipids

HOCl 诱导二酰基和烯基酰基磷脂生成溶血磷脂

• 批准号: 112537167
• 负责人: Dr. Beate Fuchs
• 金额: --
• 依托单位: Klinik und Poliklinik für Kinder- und Jugendmedizin
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/112537167?language=en
• 关键词: HOCl; induced; lysophospholipid; generation; diacyl

Inflammatory processes play extraordinarily important roles in many diseases. On the molecular level inflammatory processes are characterized by the invasion of typical inflammatory cells (such as neutrophilic granulocytes) from the blood stream into the inflamed tissue. These cells generate (beside other compounds) reactive oxygen species (ROS), for instance, HOCl. As biological samples contain always major amounts of phospholipids (PLs), their ROS-induced oxidation products possess unequivocally physiological relevance. The addition of HOCl to the unsaturated fatty acyl residues of phosphatidylcholine (PC) leads in several steps to the generation of lysophosphatidylcholine (LPC). Lysophospholipids (LPLs), particularly LPCs, are increasingly discussed as potential disease biomarkers. As PLs of different species are widely identical, LPLs would represent (in contrast to proteins) a universal biomarker. In the last funding period the influence of the positions of the double bonds with either cis or trans configuration on the reaction of PLs with HOCl were investigated and particularly the yield of LPC determined. One major aspect was also the oxidation behavior of alkenyl-PLs (plasmalogens), as these compounds are very sensitive to oxidation. A significant influence of the position of the double bond is evident. Remarkably, the trans isomers are less sensitive to oxidation than the cis-isomers. Another important result was that hydroxyl radicals (generated by the Fenton reaction) do also result in chlorohydrin generation if the reaction is performed in the presence of physiological salt concentration. The reasons for these effects are to be further investigated in more detail by modern MS and NMR methods, whereby the in vivo relevance of the identified products will be another important aspect. Beside the products of free fatty acid oxidation, the so far unavailable phospholipids are to be synthesized (by a chemoenzymatic approach) and their oxidation products have to be identified. In addition to these simplified model experiments chemical reactions of PLs will also be studied at a higher complexity level. On the one hand, the reactions between lipid oxidation products (e.g. aldehydes generated by the cleavage of the double bond) and other abundant molecules (e.g. proteins) must be studied. On the other hand, LPLs are generated in the organism not only by oxidation but as well under the influence of the enzyme phospholipase A2 (PLA2). Therefore, it must be investigated if oxidized PLs are cleaved to the same extent by PLA2 (as the non-oxidized PLs) or if the enzyme is (partially) inhibited by the presence of oxidized functional groups. Finally, it will be proven whether the in vitro detected oxidation products are also detectable in tissue slices, i.e. within a highly complex biological matrix. These investigations will be carried out by MALDI-MS-Imaging in cooperation with the University of Münster.

炎症过程在许多疾病中发挥着极其重要的作用。在分子水平上，炎症过程的特征是典型炎症细胞（例如中性粒细胞）从血流侵入发炎组织。这些细胞（除了其他化合物外）还产生活性氧 (ROS)，例如 HOCl。由于生物样品始终含有大量磷脂 (PL)，因此它们的 ROS 诱导的氧化产物具有明确的生理相关性。将 HOCl 添加到磷脂酰胆碱 (PC) 的不饱和脂肪酰基残基上，通过几个步骤导致溶血磷脂酰胆碱 (LPC) 的生成。溶血磷脂 (LPL)，特别是 LPC，作为潜在的疾病生物标志物越来越多地被讨论。由于不同物种的 PL 广泛相同，LPL 将代表（与蛋白质相反）通用生物标志物。在最后的资助期间，研究了顺式或反式构型的双键位置对 PL 与 HOCl 反应的影响，特别是确定了 LPC 的产率。其中一个主要方面是烯基-PL（缩醛磷脂）的氧化行为，因为这些化合物对氧化非常敏感。双键位置的显着影响是显而易见的。值得注意的是，反式异构体对氧化的敏感性低于顺式异构体。另一个重要的结果是，如果反应在生理盐浓度存在下进行，羟基自由基（由芬顿反应产生）也会导致氯醇的生成。这些影响的原因有待通过现代 MS 和 NMR 方法进行更详细的进一步研究，其中所鉴定产品的体内相关性将是另一个重要方面。除了游离脂肪酸氧化产物外，还需要合成迄今为止无法获得的磷脂（通过化学酶方法），并且必须鉴定它们的氧化产物。除了这些简化的模型实验之外，PL 的化学反应也将以更高的复杂性水平进行研究。一方面，必须研究脂质氧化产物（例如双键断裂产生的醛）与其他丰富分子（例如蛋白质）之间的反应。另一方面，LPL 在生物体中不仅通过氧化产生，而且还受到磷脂酶 A2 (PLA2) 的影响。因此，必须研究氧化的 PL 是否被 PLA2（与非氧化的 PL）以相同的程度裂解，或者该酶是否（部分）被氧化官能团的存在所抑制。最后，将证明体外检测到的氧化产物是否也可以在组织切片中（即在高度复杂的生物基质内）检测到。这些研究将由 MALDI-MS-Imaging 与明斯特大学合作进行。

项目成果

Interaction of plasmenylcholine with free radicals in selected model systems

• DOI: 10.1016/j.freeradbiomed.2017.02.029
• 发表时间: 2017-05-01
• 期刊: FREE RADICAL BIOLOGY AND MEDICINE
• 影响因子: 7.4
• 作者: Broniec, A.;Zadlo, A.;Sarna, T.
• 通讯作者: Sarna, T.

Stationary phase thickness determines the quality of thin-layer chromatography/matrix-assisted laser desorption and ionization mass spectra of lipids.

• DOI: 10.1016/j.ab.2014.02.002
• 发表时间: 2014-04
• 期刊: Analytical biochemistry
• 影响因子: 2.9
• 作者: Hans Griesinger;B. Fuchs;R. Süss;Katerina Matheis;M. Schulz;J. Schiller

Unexpected products of the hypochlorous acid-induced oxidation of oleic acid: A study using high performance thin-layer chromatography-electrospray ionization mass spectrometry.

• DOI: 10.1016/j.chroma.2015.11.059
• 发表时间: 2016-03
• 期刊: Journal of chromatography. A
• 作者: J. Schröter;Hans Griesinger;Eyla Reuÿ;M. Schulz;T. Riemer;Rosmarie Süÿ;J. Schiller;B. Fuchs

A simple method to identify ether lipids in spermatozoa samples by MALDI-TOF mass spectrometry

MALDI-TOF 质谱法鉴定精子样品中醚脂的简单方法

• DOI: 10.1007/s00216-013-7147-z
• 发表时间: 2013
• 期刊: Analytical and Bioanalytical Chemistry
• 影响因子: 4.3
• 作者: Nimptsch;Zschörnig;Schiller;Müller
• 通讯作者: Müller