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本文分别从微粒体和重组细胞色素P450酶系水平,研究了3-氨基苯二酰肼(luminol)化学发光在毒理学中的应用。研究结果表明luminol化学发光法不仅可用于P450酶系活力的测定,也可用来检测反应体系中由外来化合物代谢转化而产生的自由基。自由基清除剂SOD、过氧化氢酶和二甲亚砜对多种重组酶系的化学发光均有不同程度的淬灭作用,其中以SOD作用最强,其抑制率为93～100％。44种外来化合物的重组酶系化学发光的测定结果表明,已知在代谢转化过程中形成自由基的化合物,其化学发光强度明显增强,如四氯化碳、三氯甲烷、四氯乙烯等;经代谢转化后毒作用增强的化合物,其发光强度有的增强,如二硫化碳、苯、甲苯、甲基对硫磷等,有的不变或减弱,如对硫磷、苯胺、马拉硫磷等;毒作用与代谢转化关系不很密切的化合物,其发光强度与对照比较无变化或减弱,如氨基比林等,多环芳烃类和亚硝胺类致癌物的发光强度,除3-甲基胆蒽、二甲基亚硝胺外,均无明显变化。
In this paper, the application of 3-aminophthalide chemiluminescence in toxicology was studied from microsomes and recombinant cytochrome P450 enzymes. The results show that luminol chemiluminescence method can not only be used for the determination of P450 enzyme activity, but also can be used to detect the free radicals produced by metabolic transformation of foreign compounds in the reaction system. The free radical scavengers SOD, catalase and dimethyl sulfoxide have different degrees of quenching effect on chemiluminescence of various recombinant enzymes, of which SOD is the strongest and its inhibition rate is 93-100%. The results of chemiluminescence of 44 kinds of foreign compounds showed that the compounds which formed free radicals during the process of metabolic transformation have obvious chemiluminescence intensities such as carbon tetrachloride, chloroform, tetrachlorethylene and so on. After the metabolic transformation of the toxic effects of enhanced compounds, some of its luminous intensity increased, such as carbon disulfide, benzene, toluene, methyl parathion, etc., some unchanged or weakened, such as parathion, aniline, malathion, etc. ; The toxic effects and metabolic transformation not very close to the compound, its luminous intensity compared with the control no change or decrease, such as aminopyrine, polycyclic aromatic hydrocarbons and nitrosamines carcinogens luminous intensity, in addition to 3-methyl Anthracene, dimethyl nitrosamines, no significant changes.