[Methods of study on cytochrome P450 in relation to the metabolism and toxicity of alcohol and drugs].
Metabolism of xenobiotics such as alcohol and organic solvents can be divided roughly into two types: functional and conjugation reactions. Cytochrome P450 (CYP) is the primary group of enzymes involved in the former. Therefore, CYP analysis is essential for the study of the metabolism and toxicity of these chemicals. Of the isozymes, CYP2E1 is a low-km isozyme and has a high affinity for volatile hydrocarbons of low molecular mass, and is primarily involved in the metabolism of alcohol and organic solvents. There are many methods for analysis of CYPs including CYP2E1. The measurement of the metabolic rate of the target chemical in the liver is the most common. When combined with monoclonal or polyclonal antibody to CYP, contribution of the isozymes to the metabolism can be resolved, and the expression level is also identified by western blot analysis. The use of the reconstituted system of purified CYP isozymes is useful to determine the true activity (specific activity), but not common especially in that of human tissue. There are sometimes species differences in the enzymatic character of CYP isoforms, which causes difficulties in extrapolating to humans. In these case, the use of a cDNA expression system of human CYPs can cover the difficulties. The use of transgenic animals can answer the contribution of each CYP isoform in the in vivo metabolism and toxicity. The genotype of each isozyme is analyzed by PCR method, which can identify the polymorphism as well as the susceptibility to the chemicals.
Department of Hygiene, Shinshu University School of Medicine, Matsumoto, Japan.
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Nakajima
D000438:Alcohols / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D015153:Blotting, Western; D019392:Cytochrome P-450 CYP2E1 / Q000032:analysis* / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000235:genetics; D004247:DNA / Q000032:analysis; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction; D012997:Solvents; D014050:Toluene / Q000378:metabolism*
D000438:Alcohols; D012997:Solvents; D014050:Toluene; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D019392:Cytochrome P-450 CYP2E1