Borm P J, Koster A S, Frankhuijzen-Sierevogel A, Noordhoek J
Cell Biochem Funct. 1983 Oct;1(3):161-7. doi: 10.1002/cbf.290010308.
Two cell isolation procedures, i.e. a scraping/collagenase-treatment and a new vibration procedure in EDTA containing medium, were used to isolate intestinal epithelial cells. In both cell populations the metabolism of 7-ethoxycoumarin and 7-hydroxycoumarin was studied. Moreover, the time course and extent of induction of both steps in the biotransformation were investigated after oral 3-methylcholanthrene pretreatment of the rats. Twenty four hours after 3-methylcholanthrene pretreatment (20 mg kg-1) monooxygenase activity was induced about 6-fold and 2.5-fold when studied with cells of the vibratory and enzymic procedures, respectively. Control 7-ethoxycoumarin deethylase activity and 7-hydroxycoumarin glucuronidation were about the same when comparing both methods for cell-isolation. The formation of glucuronides in cells (both methods) is significantly lowered by 3-MC pretreatment, while sulphation remains unaffected. Results indicate that using enzymic treatment of mucosal scrapings, cell-populations are obtained containing relatively more differentiated (tip) cells. A number of advantages of the new (vibration) method are: better recovery, viability and reproducibility.
采用两种细胞分离方法，即刮取/胶原酶处理法和在含乙二胺四乙酸（EDTA）的培养基中进行新的振动法，来分离肠上皮细胞。对这两种细胞群体中7-乙氧基香豆素和7-羟基香豆素的代谢进行了研究。此外，在对大鼠进行口服3-甲基胆蒽预处理后，研究了生物转化过程中这两个步骤的诱导时间进程和诱导程度。在3-甲基胆蒽预处理（20毫克/千克）24小时后，当用振动法和酶法分离的细胞进行研究时，单加氧酶活性分别诱导了约6倍和2.5倍。在比较两种细胞分离方法时，对照7-乙氧基香豆素脱乙基酶活性和7-羟基香豆素葡萄糖醛酸化作用大致相同。3-甲基胆蒽预处理会使细胞（两种方法）中葡萄糖醛酸苷的形成显著降低，而硫酸化作用则不受影响。结果表明，通过对黏膜刮片进行酶处理，可以获得含有相对更多分化（顶端）细胞的细胞群体。新的（振动）方法具有许多优点：回收率更高、活力更强且可重复性更好。
