出版社：American Society for Biochemistry and Molecular Biology
摘要：We have isolated the core aldehydes (aldehydes still bound to parent molecules) of phosphatidylcholine (PC) and cholesteryl esters (CE) from copper-catalyzed peroxidation of human plasma low (LDL) and high (HDL) density lipoproteins. The aldehydes were isolated by extraction with acidified chloroform-methanol containing 2,4-dinitrophenylhydrazine. The 2,4-dinitrophenylhydrazone (DNPH) derivatives formed were resolved by reversed phase high performance liquid chromatography (HPLC) and identified by on-line quadrupole mass spectrometry (LC/MS). The major PC core aldehydes from oxidized LDL and HDL were identified as 1-palmitoyl-(1-stearoyl) 2-(9-oxononanoyl)-, 1-palmitoyl-(1-stearoyl) 2-(8-oxooctanoyl)-, and 1-palmitoyl-(1-stearoyl) 2-(5-oxovaleroyl)-sn-glycerols after phospholipase C digestion of the DNPH derivatives of the phospholipids. The major aldehydes from peroxidation of cholesteryl esters were the 9-oxononanoyl, 8-oxooctanoyl, and 5-oxovaleroyl esters of cholesterol and 7-ketocholesterol. The core aldehydes were estimated to account for a minimum of 1-2% of the consumed linoleate and arachidonate esters. A relatively smaller yield of the PC core aldehydes from LDL compared to HDL was attributed to the presence of greater amounts of phospholipases in LDL than in HDL. More comparable yields of PC core aldehydes were obtained in the presence of phenylmethylsulfonylfluoride, which inhibits phospholipases. We conclude that peroxidation of LDL and HDL results in formation of detectable amounts of cholesteryl and glycerophospholipid esters containing aldehyde functions. The yield of PC aldehydes varies with the activity of the platelet activating factor (PAF) acetyl hydrolase.
