Rodríguez-Martín A, Remesar X, Alemany M
Departament de Bioquímica i Fisiologia, Universitat de Barcelona, Spain.
Toxicology. 1993 Jun 11;80(2-3):131-9. doi: 10.1016/0300-483x(93)90176-s.
A method has been developed and tested for the measurement of anilide hydrolysing activity in rat tissues. A concentrated solution of labelled oleyl anilide in isopropanol is added to the tissue homogenates and after incubation, the chloroform/methanol extract of the samples is chromatographed on Silicagel TLC plates and the oleic acid radioactivity is measured. The activity is time-, homogenate- and temperature-dependent, the optimal pH for measurement is 8 and there is no significant spontaneous anilide degradation. In the rat, the activity is widely distributed, with highest protein specific activity in the adipose tissues. The tissue activities of a same animal are fairly well inter-correlated, with rats showing very low activity in all tissues compared with others presenting high overall activity. The levels of activity found can easily explain the fast elimination of anilides administered to rats and their scant toxic effects. Human adipose tissue samples showed a wide range of anilide hydrolase activities per gram of protein, in general lower than in rats and with some values very low. It is postulated that this lack of anilide-hydrolising capability in some humans may be related to the incidence of the toxic oil syndrome.
已开发并测试了一种用于测量大鼠组织中苯胺水解活性的方法。将异丙醇中标记的油酰苯胺浓缩溶液加入组织匀浆中,孵育后,将样品的氯仿/甲醇提取物在硅胶TLC板上进行色谱分析,并测量油酸放射性。该活性与时间、匀浆和温度有关,测量的最佳pH值为8,且不存在明显的自发苯胺降解。在大鼠中,该活性分布广泛,在脂肪组织中的蛋白质比活性最高。同一动物的组织活性之间相关性相当好,与其他总体活性高的大鼠相比,有些大鼠在所有组织中的活性都非常低。所发现的活性水平可以很容易地解释给予大鼠的苯胺的快速消除及其轻微的毒性作用。人脂肪组织样本每克蛋白质的苯胺水解酶活性范围很广,总体上低于大鼠,有些值非常低。据推测,某些人缺乏苯胺水解能力可能与有毒油综合征的发病率有关。