Tamura S, Kawata S, Okamoto M, Tarui S
Second Department of Internal Medicine, Osaka University Medical School, Japan.
Cell Tissue Res. 1988 May;252(2):397-401. doi: 10.1007/BF00214382.
Immunohistochemical localization of cytochrome P-450 in the colonic mucosa of 3-methylcholanthrene-pretreated and untreated rats was studied by indirect fluorescent antibody staining technique. A polyclonal antibody for cytochrome P-450MC purified from hepatic microsomes of 3-methylcholanthrene-pretreated rats was used for this experiment. A strong immunofluorescence was found to be localized in the cytoplasm of the surface epithelium of the mucosa in the colon of 3-methylcholanthrene-pretreated rats. A faint immunofluorescence was also observed in the epithelium of untreated rats. 7-Ethoxycoumarin O-deethylase activity of colonic microsomes was significantly enhanced by 3-methylcholanthrene-pretreatment in parallel with an increase in the intensity of immunostaining for cytochrome P-450MC in Western blotting analysis. This is the first report on the localization of cytochrome P-450 in the colonic mucosa.
采用间接荧光抗体染色技术,研究了经3-甲基胆蒽预处理和未经处理的大鼠结肠黏膜中细胞色素P-450的免疫组织化学定位。本实验使用了从经3-甲基胆蒽预处理的大鼠肝微粒体中纯化得到的细胞色素P-450MC多克隆抗体。在经3-甲基胆蒽预处理的大鼠结肠黏膜表面上皮细胞的细胞质中发现了强烈的免疫荧光。在未经处理的大鼠上皮细胞中也观察到微弱的免疫荧光。在蛋白质免疫印迹分析中,3-甲基胆蒽预处理显著增强了结肠微粒体的7-乙氧基香豆素O-脱乙基酶活性,同时细胞色素P-450MC免疫染色强度增加。这是关于细胞色素P-450在结肠黏膜中定位的首次报道。