title: Hepatic transcriptome and proteome responses against diethyl maleate-induced glutathione depletion in the rat.
abstract: Hepatic transcriptome and proteome responses against glutathione depletion were investigated by Affymetrix GeneChip Microarray and 2-dimensional gel electrophoresis (2D-DIGE), followed by MALDI-TOF-MS analysis and utilizing a glutathione-depleted rat model treated with diethyl maleate (DEM). Hepatic glutathione content decreased to 1.29 μmol/g liver (25.5% compared to control) after DEM treatment, and there were no apparent hepatotoxic signs estimated by blood chemistry examinations. A total of 247 and 213 annotated gene probe sets exhibited greater than twofold up- and down-regulation compared with controls, respectively. The up-regulated gene list contained a number of glutathione depletion-responsive genes reported previously, such as Trib3, Srxn1, Myc, Asns, Igfbp1, Txnrd1, or Hmox1, suggesting that these genes are robust mRNA biomarkers for evaluating hepatic glutathione depletion. In the 2D-DIGE analysis, proteins for a total of 361 spots were identified by MALDI-TOF-MS analysis. Of the identified proteins, 5 and 14 proteins showed up- and down-regulation, respectively. Some proteins exhibited differential expression in the protein level but not in the mRNA level, including L-FABP, MAWDBP, aldo-keto reductase family 1 member A1, catalase and ATP synthase subunit beta, suggesting that these proteins would be potential protein biomarkers for evaluating glutathione depletion. Moreover, up-regulation of FABP1 protein along with up-regulation of PPARα-regulated gene transcripts (i.e., Acot2 and Acot4) is indicative of PPARα activation, which may contribute to hepatocellular protection against glutathione depletion-induced oxidative stress. The up-regulation of L-FABP1 was detected by proteome data but not by transcriptome data, demonstrating the advantage of utilizing transcriptomics and proteomics combination to investigate glutathione depletion-induced molecular dynamics.
authors.affiliation: Medicinal Safety Research Laboratories, Daiichi Sankyo Co., Ltd., Shizuoka, Japan.
authors.forename: Shusuke
authors.initials: S
authors.lastname: Yamauchi
authors.forename: Naoki
authors.initials: N
authors.lastname: Kiyosawa
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Ando
authors.forename: Kyoko
authors.initials: K
authors.lastname: Watanabe
authors.forename: Noriyo
authors.initials: N
authors.lastname: Niino
authors.forename: Kazumi
authors.initials: K
authors.lastname: Ito
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamoto
authors.forename: Sunao
authors.initials: S
authors.lastname: Manabe
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sanbuissho
mesh_terms: D000818:Animals; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D005978:Glutathione / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008111:Liver Function Tests; D008297:Male; D008298:Maleates / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D051381:Rats; D011916:Rats, Inbred F344; D043922:Toxicogenetics / Q000379:methods; D059467:Transcriptome / Q000187:drug effects*; D015854:Up-Regulation
chemical_list: D008298:Maleates; D020543:Proteome; C014476:diethyl maleate; D005978:Glutathione