Lev R, Griffiths W C
Gastroenterology. 1982 Jun;82(6):1427-35.
Colonic and intestinal alkaline phosphatase were studied histoenzymatically and biochemically in normal mucosae from eight hemicolectomy specimens. Histochemically, the most intense reaction was found in the small intestine, followed by vascular endothelium and colonic epithelium, respectively. The reaction was stronger in frozen sections than in acetone-fixed specimens, weakest in formalin-fixed tissues, and best when cold fixatives and low melting point paraffin were used. In the small intestine, the reaction was most marked on the villus tips. In the colon, the crypt necks showed the strongest reaction although good staining of luminal epithelium was found with more prolonged incubation. The addition of phenylalanine to the substrate medium resulted in selective inhibition of epithelial staining, whereas levamisole selectively inhibited the vascular reaction. On cellulose acetate electrophoresis, colonic and intestinal alkaline phosphatase showed slightly different rates of migration suggesting the presence of different isoenzymes. Quantitative analysis indicated that the intestinal enzyme expressed in U/g wet tissue was 7.7 times the colonic enzyme in amount. It is concluded that despite its relatively low biochemical activity, alkaline phosphatase can be demonstrated histochemically in colonic epithelium if tissues are processed at low temperatures and if the appropriate fixatives and adequate times are used.
对取自8个半结肠切除术标本的正常黏膜组织进行了组织酶学和生物化学研究,以观察结肠和小肠碱性磷酸酶。组织化学研究发现,小肠的反应最为强烈,其次是血管内皮和结肠上皮。冷冻切片的反应强于丙酮固定标本,福尔马林固定组织的反应最弱,使用冷固定剂和低熔点石蜡时染色效果最佳。在小肠中,绒毛尖端的反应最为明显。在结肠中,隐窝颈部的反应最强,不过延长孵育时间可使腔面上皮获得良好染色。向底物培养基中添加苯丙氨酸可选择性抑制上皮染色,而左旋咪唑则选择性抑制血管反应。在醋酸纤维素电泳中,结肠和小肠碱性磷酸酶的迁移速率略有不同,提示存在不同的同工酶。定量分析表明,以每克湿组织中的单位数表示,小肠酶的含量是结肠酶的7.7倍。研究得出结论,尽管碱性磷酸酶的生化活性相对较低,但如果在低温下处理组织,并使用适当的固定剂和足够的时间,仍可在结肠上皮中通过组织化学方法显示该酶。