大鼠肝脏缺血再灌注产生的过氧化氢磷脂酰胆碱的化学发光-HPLC测定法

Chemiluminescence-HPLC assay of phosphatidylcholine hydroperoxide generated by ischemia-reperfusion in the liver of rats.

作者信息

Takayama F, Egashira T, Kudo Y, Yamanaka Y

机构信息

Department of Pharmacology, Oita Medical University, Japan.

出版信息

Biochem Pharmacol. 1992 Dec 15;44(12):2412-4. doi: 10.1016/0006-2952(92)90688-f.

DOI:10.1016/0006-2952(92)90688-f

PMID:1472107

Abstract

To determine cellular damage due to "oxidative stress", we developed a sensitive and specific quantitative assay for phosphatidylcholine hydroperoxide (PCOOH) by coupling HPLC with detection of chemiluminescence (CL). The qualitative and quantitative detection limits of PCOOH by this assay were 0.5 and 2 pmol (based on active oxygen from hydroperoxide). Using this CL-HPLC method, we determined PCOOH levels caused by ischemia-reperfusion in rat livers. The PCOOH levels in livers of control, sham-operated and operated rats with only ischemic treatment were approximately 2 nmol/g wet liver weight. The PCOOH level and several serum parameters of liver injury increased with an increase in the duration of ischemia, and also increased in proportion to the duration of reperfusion. The determination of PCOOH in liver caused by ischemia-reperfusion could be a useful method for investigating liver damage induced by free radicals.

摘要

为了确定“氧化应激”导致的细胞损伤，我们通过将高效液相色谱（HPLC）与化学发光（CL）检测相结合，开发了一种灵敏且特异的磷脂酰胆碱氢过氧化物（PCOOH）定量检测方法。该检测方法对PCOOH的定性和定量检测限分别为0.5和2皮摩尔（基于氢过氧化物产生的活性氧）。使用这种CL-HPLC方法，我们测定了大鼠肝脏缺血再灌注引起的PCOOH水平。对照组、假手术组和仅接受缺血处理的手术组大鼠肝脏中的PCOOH水平约为2纳摩尔/克湿肝重。肝脏损伤的PCOOH水平和几个血清参数随着缺血时间的延长而增加，并且与再灌注时间成正比增加。测定缺血再灌注引起的肝脏中的PCOOH可能是研究自由基诱导的肝损伤的一种有用方法。