Camp R, Greaves M W
Biochem J. 1980 Jan 15;186(1):153-60. doi: 10.1042/bj1860153.
The activities of NAD+-dependent 15-hydroxy prostaglandin dehydrogenase in soluble fractions of rat skin and lung were compared by using a radiochemical assay method. Tritiated prostaglandin F2 alpha was incubated with NAD+ and 120,000 g supernatant of tissue homogenate. Extracted prostaglandin substrate and reaction products were separated by t.l.c. and quantitatively determined by liquid-scintillation counting. With skin 120,000 g supernatant, 10 mM-NAD+ and an incubation time of 15 min, the mean Vmax. was 5.5 nmol of prostaglandin F2 alpha converted/s per litre of reaction mixture. With lung 120,000 g supernatant, 60 mM-NAD+ and an incubation time of 5 min, the mean Vmax. was 26.9 nmol/s per litre, demonstrating 5-fold greater dehydrogenase activity in lung per unit wet weight of tissue. However, the total wet weight of skin was about 23 times that of lung, on dissection of individual rats, indicating that the entire skin may contain 4.5 times the total 15-hydroxy prostaglandin dehydrogenase activity of the lungs. Skin may thus be an important organ of prostaglandin catabolism.
采用放射化学分析法比较了大鼠皮肤和肺可溶性组分中NAD⁺依赖性15-羟基前列腺素脱氢酶的活性。将氚标记的前列腺素F2α与NAD⁺及组织匀浆120,000g上清液一起孵育。提取的前列腺素底物和反应产物通过薄层层析分离,并用液体闪烁计数法定量测定。对于皮肤120,000g上清液、10mM-NAD⁺及15分钟的孵育时间,平均Vmax为每升反应混合物每秒转化5.5nmol前列腺素F2α。对于肺120,000g上清液、60mM-NAD⁺及5分钟的孵育时间,平均Vmax为每升每秒26.9nmol,表明每单位组织湿重肺中的脱氢酶活性高5倍。然而,在解剖个体大鼠时,皮肤的总湿重约为肺的23倍,这表明整个皮肤可能含有肺中15-羟基前列腺素脱氢酶总活性的4.5倍。因此,皮肤可能是前列腺素分解代谢的重要器官。