Liquid chromatography mass spectrometry-based profiling of phosphatidylcholine and phosphatidylethanolamine in the plasma and liver of acetaminophen-induced liver injured mice

Ya-Nan Ming,Xiao-Lin Wang,Chun-Min Li,Xiao-Lin Liu,Xiao-Bo Li,Si-Cong Ma,Yi-Min Mao,Jing-Yi Zhang,Zheng-Yang Wang

doi:10.1186/s12944-017-0540-4

Abstract

BackgroundAcetaminophen (APAP) overdose is one of the most common causes of acute liver failure in many countries. The aim of the study was to describe the profiling of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in the plasma and liver of Acetaminophen -induced liver injured mice.MethodsA time course study was carried out using C57BL/6 mice after intraperitoneal administration of 300 mg/kg Acetaminophen 1 h, 3 h, 6 h, 12 h and 24 h. A high-throughput liquid chromatography mass spectrometry (LC-MS) lipidomic method was utilized to detect phosphatidylcholine and phosphatidylethanolamine species in the plasma and liver. The expressions of phosphatidylcholine and phosphatidylethanolamine metabolism related genes in liver were detected by quantitative Reverse transcription polymerase chain reaction (qRT-PCR) and Western-blot.ResultsFollowing Acetaminophen treatment, the content of many PC and PE species in plasma increased from 1 h time point, peaked at 3 h or 6 h, and tended to return to baseline at 24 h time point. The relative contents of almost all PC species in liver decreased from 1 h, appeared to be lowest at 6 h, and then return to normality at 24 h, which might be partly explained by the suppression of phospholipases mRNA expressions and the induction of choline kinase (Chka) expression. Inconsistent with PC profile, the relative contents of many PE species in liver increased upon Acetaminophen treatment, which might be caused by the down-regulation of phosphatidylethanolamine N-methyltransferase (Pemt).ConclusionsAcetaminophen overdose induced dramatic change of many PC and PE species in plasma and liver, which might be caused by damaging hepatocytes and interfering the phospholipid metabolism in Acetaminophen -injured liver.

Highlights

Acetaminophen (APAP) overdose is one of the most common causes of acute liver failure in many countries
The prominent morphological damage started at 3 h, deteriorated with time, and attenuated at 24 h time point (Fig.1d)
Phosphatidylcholine and phosphatidylethanolamine profiling in mice plasma analyzed by liquid chromatography mass spectrometry (LC-MS) The relative concentrations of 57 PC/LPC species and 18 PE/LPE species in plasma were simultaneously determined using LC-MS method

Summary

Introduction

Acetaminophen (APAP) overdose is one of the most common causes of acute liver failure in many countries. As a major site for drug metabolism and elimination, the liver is susceptible to drug toxicity. Drug-induced liver injury (DILI) is a significant clinical problem and a challenge for drug development worldwide. APAP overdose has become one of the most common causes of acute liver failure in many countries [1]. APAPinduced liver injury is the most frequent drug hepatotoxicity and the most used experimental model of DILI. The mechanism of APAP-induced liver injury is complicated and not fully understood. The accumulation of Nacetyl-p-benzoquinone imine (NAPQI), the reactive and toxic metabolite of APAP, is considered as the main cause of liver injury induced by overdose of APAP. In mouse APAP-induced models and in human, the reaction of NAPQI with protein sulfhydryl groups of cysteine might trigger mitochondrial damage, oxidative stress, cjun N-terminal kinase (JNK) activation, the nuclear DNA fragmentation and cell death [2,3,4]

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