title: An improved in vitro method for screening toxin and medicine targeting CYP2E1.
abstract: The cytochrome P450 enzyme 2E1 (CYP2E1) presents in both microsome and mitochondrion, which influences the metabolism of many xenobiotics. The mice active liver homogenate was prepared for the medicinal incubation and mitochondrion was extracted for chemical screening targeting CYP2E1 enzyme. Representative CYP2E1 inducers (ethanol and pyrazole) and inhibitors (diallyldisulfide and kaempferol) were applied to evaluate the effectiveness of homogenate-mitochondrial system. In parallel, the in-vitro microsomal method targeting CYP2E1 was also operated for comparison. The results showed that in homogenate-mitochondrial method, the protein level and activity of CYP2E1 were increased by ethanol and pyrazole; reduced by diallyldisulfide and kaempferol, and this homogenate-mitochondrial method is convenient with good repeatability and reproducibility in screening chemicals targeting CYP2E1, especially for the inducers. Thus, the homogenate-mitochondrial method might be effective in screening both CYP2E1 inhibitor and inducer.
authors.affiliation: Chinese-German Joint Laboratory for Natural Product Research, Qinling-Bashan Mountains Bioresources Comprehensive Development C.I.C., College of Biological Science and Engineering, Shaanxi Sci-tech University, Hanzhong, Shaanxi 723000, China; College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
authors.forename: Zhaoxiang
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Chinese-German Joint Laboratory for Natural Product Research, Qinling-Bashan Mountains Bioresources Comprehensive Development C.I.C., College of Biological Science and Engineering, Shaanxi Sci-tech University, Hanzhong, Shaanxi 723000, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.affiliation: Chinese-German Joint Laboratory for Natural Product Research, Qinling-Bashan Mountains Bioresources Comprehensive Development C.I.C., College of Biological Science and Engineering, Shaanxi Sci-tech University, Hanzhong, Shaanxi 723000, China; College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China. Electronic address: zhang.xy@nwsuaf.edu.cn.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000818:Animals; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism*; D065700:Cytochrome P-450 CYP2E1 Inducers / Q000494:pharmacology*; D065691:Cytochrome P-450 CYP2E1 Inhibitors / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D051379:Mice; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism; D015203:Reproducibility of Results; D014118:Toxins, Biological / Q000633:toxicity
chemical_list: D065700:Cytochrome P-450 CYP2E1 Inducers; D065691:Cytochrome P-450 CYP2E1 Inhibitors; D014118:Toxins, Biological; D019392:Cytochrome P-450 CYP2E1
keywords: Cytochrome 2E1; Drug screening; Liver homogenate; Microsome; Mitochondrion