ABSTRACT
OBJECTIVE To explore the effect of aqueous extract of Zheng Chaihu Yin(ZCH)on paracetamol(acetaminophen,APAP)-induced hepatotoxicity. METHODS Male ICR mice were divided into three scenarios randomly:the single treatment dose of ZCH,multiple treatment or pretreatment dose of ZCH. Each scenario had a up control group and an APAP model group,while single treatment dose of ZCH group had a ZCH group at the same time. The dose of APAP and ZCH was 500 mg·kg-1 and 36 g · kg- 1,respectively. 24 h after the last administration,plasma and liver samples were prepared. Ultra- performance liquid chromatography/quadrupole- time- of- flight mass spectrometry (UPLC-Q-TOF-MS)based metabolomics profiling was used to examine changes in plasma after expo?sure to ZCH,APAP or co-exposure to ZCH and APAP. Glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminas (GOT) values were determined by a biochemical auto analyzer in plasma. Histopathologic changes in the liver were observed and the area was calculated after HE staining. The data were analyzed with SPSS16.0 statistical software and the results were compared with the test between the two groups to find biomarkers. Also,SIMCA software was used for partial least squares-discriminant analysis (PLS-DA) pattern recognition. RESULTS Compared to control group, APAP dosing alone caused an increase in plasma transaminases and alterations in multiple metabolic pathways. Compared to APAP group,decrease in plasma transaminases was noted when ZCH was administered after or prior to APAP. Histopathologic results showed that in the single treatment group, multiple treatment group and pretreatment group,ZCH could alleviate the liver damage induced by APAP from (32.3 ± 12.0)% to (14.2 ± 9.9)%,(8.6 ± 7.9)% to (5.2 ± 1.7)% and (32.5 ± 10.0)% to (5.2 ± 6.4)%(P<0.05). Similarly,the PLS-DA of the LC-MS data showed that the groups dosed with APAP alone were the most distinct from controls,while animals dosed with ZCH prior to or after APAP treatment were located near control group. Metabolic spectrum results showed that ZCH could restore the changes in endogenous substances including lipid metabolism,amino acid metabolism,sugar metabolism and energy metabolism induced by APAP to normal. CONCLUSION ZCH water-extraction plays major roles in the regulation of metabolism on APAP-induced liver injury. These studies demonstrate that UPLC-Q-TOF-MS-based metabolomic analysis can be sensitively and accurately predict the initiation and progres?sion of liver injury and greatly contribute to a better understanding of the hepatoprotective effects of ZCH in a clinical environment.