Bhagwat S V, Boyd M R, Ravindranath V
Department of Neurochemistry, National Institute of Mental Health and Neuro Sciences, Bangalore, India.
Arch Biochem Biophys. 1995 Jun 20;320(1):73-83. doi: 10.1006/abbi.1995.1344.
The capability of rat brain mitochondria to metabolize a variety of xenobiotics was examined. The presence of cytochrome P450 (P450) and associated monooxygenase activities were estimated in isolated rat brain mitochondria and compared with the corresponding activities in microsomes. Total P450 content in brain mitochondria from naive rats was twice that of the corresponding microsomal level. The ability of brain mitochondria to metabolize the potent carcinogen N-nitrosodimethylamine was more than twofold that of the corresponding microsomal activity, while the 7-ethoxycoumarin-O-deethylase activity was significantly lower in mitochondria. Immunoblot experiments using antisera to purified rat liver microsomal P450s, namely P450 (2B1/2B2), P4501A1, and P4502E1, and purified phenobarbital-inducible rat brain P450, revealed the presence of immunoreactive bands in isolated brain mitochondria. These various antibodies to P450 inhibited the brain mitochondrial monooxygenase activities to significant, though varying extent. The addition of antiserum to microsomal NADPH cytochrome P450 reductase did not affect the mitochondrial P450 associated monooxygenase activities, although it completely inhibited the corresponding microsomal activities. Chronic ethanol administration resulted in twofold induction of total P450 content and the monooxygenase activities known to be mediated by P4502E1, such as N-nitrosodimethylamine-N-demethylase and p-nitrophenol hydroxylase in brain mitochondria. Pretreatment of animals with phenobarbital resulted in the induction of aminopyrine N-demethylase activity in brain mitochondria. The study demonstrates the presence of multiple forms of P450 in the rat brain mitochondria, their inducibility, and their capability to metabolize xenobiotics.
研究了大鼠脑线粒体代谢多种外源化合物的能力。在分离的大鼠脑线粒体中评估了细胞色素P450(P450)的存在及相关单加氧酶活性,并与微粒体中的相应活性进行比较。未接触过药物的大鼠脑线粒体中的总P450含量是相应微粒体水平的两倍。脑线粒体代谢强致癌物N-亚硝基二甲胺的能力是相应微粒体活性的两倍多,而7-乙氧基香豆素-O-脱乙基酶活性在线粒体中显著较低。使用针对纯化的大鼠肝微粒体P450(即P450(2B1/2B2)、P4501A1和P4502E1)以及纯化的苯巴比妥诱导的大鼠脑P450的抗血清进行免疫印迹实验,结果显示在分离的脑线粒体中存在免疫反应条带。这些针对P450的各种抗体对脑线粒体单加氧酶活性有显著抑制作用,尽管抑制程度有所不同。添加针对微粒体NADPH细胞色素P450还原酶的抗血清并不影响线粒体中与P450相关的单加氧酶活性,尽管它完全抑制了相应的微粒体活性。长期给予乙醇导致总P450含量增加两倍,以及脑线粒体中已知由P4502E1介导的单加氧酶活性增加,如N-亚硝基二甲胺-N-脱甲基酶和对硝基苯酚羟化酶。用苯巴比妥预处理动物导致脑线粒体中氨基比林N-脱甲基酶活性增加。该研究证明大鼠脑线粒体中存在多种形式的P450、它们的可诱导性以及代谢外源化合物的能力。