title: Proteomic analysis for the impact of hypercholesterolemia on expressions of hepatic drug transporters and metabolizing enzymes.
abstract: 1. Our objective is to investigate the alterations of hepatic drug transporters and metabolizing enzymes in hypercholesterolemia. Male Sprague-Dawley rats were fed high-cholesterol chows for 8 weeks to induce hypercholesterolemia. Protein levels of hepatic drug transporters and metabolizing enzymes were analyzed by iTRAQ labeling coupled with LC TRIPLE-TOF. 2. Total 239 differentially expressed proteins were identified using proteomic analysis. Among those, protein levels of hepatic drug transporters (MRP2, ABCD3, OAT2, SLC25A12, SCL38A3, SLC2A2 and SLC25A5) and metabolizing enzymes (CYP2B3, CYP2C7, CYP2C11, CYP2C13, CYP4A2 and UGT2B) were markedly reduced, but the levels of CYP2C6 and CYP2E1 were increased in hypercholesterolemia group compared to control. Decreased expressions of drug transporters MRP2 and OAT2 were further confirmed by real time quantitative PCR (RT-qPCR) and western blot. 3. Ingenuity pathway analysis revealed that these differentially expressed proteins were regulated by various signaling pathways including nuclear receptors and inflammatory cytokines. One of the nuclear receptor candidates, liver X receptor alpha (LXRα), was further validated by RT-qPCR and western blot. Additionally, LXRα agonist T0901317 rescued the reduced expressions of MRP2 and OAT2 in HepG2 cells in hypercholesterolemic serum treatment. 4. Our present results indicated that hypercholesterolemia affected the expressions of various drug transporters and metabolizing enzymes in liver via nuclear receptors pathway. Especially, decreased function of LXRα contributes to the reduced expressions of MRP2 and OAT2.
authors.affiliation: a Institute of Life Science, Chongqing Medical University , Chongqing , P.R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: a Institute of Life Science, Chongqing Medical University , Chongqing , P.R. China.
authors.forename: Qiang-Hong
authors.initials: QH
authors.lastname: Pu
authors.affiliation: a Institute of Life Science, Chongqing Medical University , Chongqing , P.R. China.
authors.forename: Ming-Jun
authors.initials: MJ
authors.lastname: Wu
authors.affiliation: a Institute of Life Science, Chongqing Medical University , Chongqing , P.R. China.
authors.forename: Chao
authors.initials: C
authors.lastname: Yu
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D033742:Adenine Nucleotide Translocator 2 / Q000378:metabolism; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D001692:Biological Transport; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D000072467:Cytochrome P450 Family 2 / Q000378:metabolism; D051273:Glucose Transporter Type 2 / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000378:metabolism; D006937:Hypercholesterolemia / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism; D015778:Minor Histocompatibility Antigens / Q000378:metabolism; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism; D029362:Organic Anion Transporters, Sodium-Independent / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D039181:Steroid 16-alpha-Hydroxylase / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; C489272:Abcd3 protein, rat; D033742:Adenine Nucleotide Translocator 2; D051273:Glucose Transporter Type 2; D008840:Microfilament Proteins; D015778:Minor Histocompatibility Antigens; D033681:Mitochondrial Membrane Transport Proteins; D029362:Organic Anion Transporters, Sodium-Independent; D020543:Proteome; C088800:Slc22a7 protein, rat; C491957:Slc25a12 protein, rat; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; C450279:CYP2C11 protein, rat; C089142:Cyp2b3 protein, rat; C113764:Cyp2c13 protein, rat; C493085:Cyp2c7 protein, rat; D000072467:Cytochrome P450 Family 2; D039181:Steroid 16-alpha-Hydroxylase; C465388:cytochrome P-450 CYP4A2 (rat); D014453:Glucuronosyltransferase; C583026:glucuronosyltransferase 2B, rat
keywords: Drug metabolizing enzymes; drug transporters; hypercholesterolemia; liver X receptor