Burke M D, Murray G I, Lees G M
Biochem J. 1983 Apr 15;212(1):15-24. doi: 10.1042/bj2120015.
A fluorescence-microscopic method has been developed for measurement of the intracellular kinetics of the cytochrome P-450 reaction, ethoxyresorufin O-de-ethylation, in individual hepatocytes in unfixed non-frozen liver sections obtained from control or 3-methylcholanthrene-pretreated mice. De-ethylation was enhanced in the presence of salicylamide, but inhibited in the presence of alpha-naphthoflavone, A total of 21 reaction rate curves (fluorescence of the metabolite, resorufin, versus time) were constructed for different individual hepatocytes and groups of two or three hepatocytes, in a total of 12 sections from four control and eight 3-methylcholanthrene-induced mice. No two reaction curves were identical, but the curves were classified, by similarity of curve shape and fluorescence intensity, into three control categories and four 3-methylcholanthrene-induced categories. It was considered that the data represented a total of seven types of hepatocytes, which differed in their ethoxyresorufin de-ethylase reaction characteristics. Hepatocytes in the sections from 3-methylcholanthrene-pretreated mice showed higher de-ethylase activities than the hepatocytes from control mice, but the increase in activity covered a wide range of values, from 3-fold to 17-fold higher than the mean control value, suggesting that 3-methylcholanthrene did not induce ethoxyresorufin de-ethylation to the same extent in all hepatocytes.
已开发出一种荧光显微镜方法，用于测量从对照或经3 - 甲基胆蒽预处理的小鼠获得的未固定、未冷冻肝脏切片中单个肝细胞内细胞色素P - 450反应（乙氧萘胺O - 脱乙基反应）的动力学。在水杨酰胺存在下脱乙基作用增强，但在α - 萘黄酮存在下受到抑制。总共为不同的单个肝细胞以及两个或三个肝细胞组成的组构建了21条反应速率曲线（代谢物试卤灵的荧光与时间的关系），这些肝细胞来自4只对照小鼠和8只经3 - 甲基胆蒽诱导的小鼠的总共12个切片。没有两条反应曲线是相同的，但根据曲线形状和荧光强度的相似性，这些曲线被分为三个对照类别和四个3 - 甲基胆蒽诱导类别。据认为，这些数据代表了总共七种类型的肝细胞，它们的乙氧萘胺脱乙基酶反应特性不同。来自经3 - 甲基胆蒽预处理小鼠的切片中的肝细胞显示出比对照小鼠肝细胞更高的脱乙基酶活性，但活性增加的范围很广，比对照平均值高3倍至17倍，这表明3 - 甲基胆蒽并非在所有肝细胞中同等程度地诱导乙氧萘胺脱乙基反应。
