D'Agostino L, Daniele B, Pignata S, D'Argenio G, Mazzacca G
Cattedra di Gastroenterologia, 2a Facolta' di Medicina, Universita' di Napoli, Italy.
Biochem Pharmacol. 1989 Jan 1;38(1):47-9. doi: 10.1016/0006-2952(89)90147-0.
Heparin releases diamine oxidase (DAO) of enterocytic origin from binding sites located on small bowel microvascular endothelium. In the villus tip enterocytes the enzyme is found in organelles (about 60%) and in cytosol (about 40%), while a negligible activity is present in the brush border. In this study we assessed the changes in DAO distribution into the enterocytes induced by a high dose of intraperitoneal heparin (1000 IU) in the rat, by assaying DAO activity on subcellular fractions obtained from ileal mucosa homogenate. Heparin injection induced a marked reduction of enzyme activity in the S2 fraction (cytosol): after 30 min less than 20% of DAO activity is still found and only 8% after 150 min. In the P1 fraction (organelles) DAO activity significantly decreased only after 60 min and a further consistent reduction was recorded after 150 min. Recovery of DAO activity was complete 4 days after the injection, though it was already clearly evident in the first 2 days. These results indicate that enterocytic DAO is distributed in two different compartments: DAO located in the cytosol is quickly released by heparin, while the organelles-linked enzyme is more slowly released. The finding that recovery in DAO activity happens earlier in the P1 fraction suggests that the enzyme supplies the cytosol after being synthesized in the enterocyte organelles.
肝素可从小肠微血管内皮细胞上的结合位点释放肠细胞来源的二胺氧化酶(DAO)。在绒毛顶端的肠细胞中,该酶存在于细胞器(约60%)和胞质溶胶(约40%)中,而刷状缘中的活性可忽略不计。在本研究中,我们通过检测从回肠黏膜匀浆获得的亚细胞组分中的DAO活性,评估了高剂量腹腔注射肝素(1000 IU)对大鼠肠细胞中DAO分布的影响。注射肝素后,S2组分(胞质溶胶)中的酶活性显著降低:30分钟后,仍可检测到的DAO活性不到20%,150分钟后仅为8%。在P1组分(细胞器)中,DAO活性仅在60分钟后显著降低,150分钟后进一步显著降低。注射后4天,DAO活性完全恢复,尽管在最初2天就已明显可见。这些结果表明,肠细胞中的DAO分布在两个不同的区室:位于胞质溶胶中的DAO可被肝素快速释放,而与细胞器相连的酶释放较慢。P1组分中DAO活性恢复较早这一发现表明,该酶在肠细胞细胞器中合成后供应至胞质溶胶。