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Fast Evaluation of Oxidative DNA Damage by Liquid Chromatography-Electrospray Tandem Mass Spectrometry Coupled With precision-cut Rat liver slices

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Biomedical and Environmental Sciences 2007年第5期20卷 386-391页

作者： JIANG YUE PENG WANG YING-HUI LIU JUN-YU WU JIE CHEN REN-XIU PENG Department of Pharmacology College of Basic Medicine Wuhan University Wuhan 430071 Hubei China Inspection ＆ Quarantine Technical Center Hubei Entry-exit Inspection and Quarantine Bureau Wuhan 430022 Hubei China

Objective To establish a fast and sensitive method for the detection of 8-hydroxy-2’-deoxyguanosine （8-OHdG） in precision-cut rat liver slices by HPLC-MS/MS and to investigate isoniazid （INH） -induced oxidative DNA damage. Methods precision-cut liver slices （300 μm） were prepared from male rats, and incubated with INH （0.018 mol/L） for 2 h after 1 h preincubation. DNA in the slices was extracted and digested into free nucleosides at 37℃ . The samples were injected into HPLC-MS/MS after the proteins were removed. The level of oxidative DNA damage was estimated using the ratio of 8-OHdG to deoxyguanosine （dG）. Results The limit of detection of 8-OHdG was 1 ng/mL （S/N=3） and the intra-assay relative standard variation was 3.38% when one transition 284.3/168.4 was used as a quantifier and another two transitions 284.3/140.2, 306.1/190.2 as qualifiers. 8-OHdG and dG were well separated, as indicated by elution at 10.02 and 7.37 min, respectively. INH significantly increased the ratio of 8-OHdG to dG in rat liver slices （P〈0.05）. Conclusion 8-OHdG in precision-cut liver slices could be sensitively determined by HPLC-MS/MS. HPLC-MS/MS coupled with precision-cut tissue slices is a fast and reliable analytical technique to evaluate oxidative DNA damage of target tissues caused by procarcinogens and cytotoxins.

关键词： Isoniazid 8-Hydroxy-2'-deoxyguanosine HPLC-MS/MS precision-cut liver slices

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Protection of sodium ferulate on ethanol-induced hepatotoxicity in rat precision-cut liver slices and its mechanism

Protection of sodium ferulate on ethanol-induced hepatotoxic...

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第十五届国际药理学大会

作者： Yu GUO Xiao-qian WU Chun ZHANG Hui WANG Zhang - xiu LIAO Yong WU Department of Pharmacology Basic Medical School of Wuhan University Wuhan 430071 China

＜正＞ To investigate the protection and mechanism of sodium ferulate （SF） on ethanol- induced toxicity in rat precision - cut liver slices （PCLS）. PCLS were incubated with SF （0.5 - 2mM） either co - treated with ethanol or its metabolite ac etaldehyde. The releases of glutathione S - transferase （GST） and lactate dehy drogenase （LDH）. the activities of aniline hydroxylase （ANH） and alcohol de hydrogenase （ADH） and the content of malondialdehyde （MDA） were monitored. Meanwhile, the contents of hydroxyproline （Hyp）, transform grow factor- β1（ TGF- β1） and α - smooth muscle actin （α- SMA） were detected. The results showed that SF reduce the leakage of enzymes, degrade lipid peroxidation. and turn ANH and ADH to the normal level. Prominent inhibition of HSC activation was achieved with SF against acetaldehyde - induced increase of α - SMA and TGF -β1. and Hyp content showed a decrease tendency. The results demonstrated that SF exerts protective effects on ethanol - induced hepatotoxicity. which attribute to the modification of ethanol - metabolizing pathway and inhibition of HSC activation.

关键词： sodium ferulate precision-cut liver slices ethanol acetaldehyde.

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Application of precision-cut Rat liver Slice to Study the Influence of Monocrotaline, Tussilago farfara Alkaloids on the Expression of Cytochrome P450 Enzymes

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Health 2016年第4期8卷 370-379页

作者： Hailin Wang Lianqiang Hui Chun Li Ting Liu Chang’an Yu Chunyu Cao Ran Hao Yi Zhang Institute of Chinese Materia Medica China Academy of Chinese Medical Sciences Beijing China China-Japanese Friendship Hospital Beijing China

precision-cut liver slice has been successfully used to study the mechanism of drug-induced hepatotoxicity, the prediction of liver toxicity, the discovery of early hepatic toxicity biomarker and the metabolism of drug in liver. We detected the expression of CYP3A4, CYP2B1 + CYP2B2 and CYP2E1 in precision-cut liver slice after co-cultured with monocrotaline or Tussilago farfara alkaloids to investigate the hepatotoxicity mechanism of those drugs. After co-culturing with monocrotaline or Tussilago farfara alkaloids for 6 hours, the expression of CYP3A4 in the microsome of precision-cut liver slices was detected by Western blot, and the expressions of CYP2B1 + CYP2B2 and CYP2E1 were detected by immunofluorescence. The results showed that monocrotaline induced the expression of CYP3A4 and CYP2B1 + CYP2B2, and Tussilago farfara alkaloids obviously up-regulated the expression of CYP2E1 and CYP3A4. Thus, we conclude that the up-regulation of CYP3A4, CYP2B1 + CYP2B2 and CYP2E1 may be one of the toxic mechanisms of liver injury of those drugs.

关键词： precision-cut liver slices Monocrotaline Tussilago farfara Alkaloids Hepatotoxity Cytochrome P450 Enzymes

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