Young H M, O'Brien A J, Furness J B, Ciampoli D, Hardwick J P, McCabe T J, Narayanasami R, Masters B S, Tracey W R
Department of Anatomy and Cell Biology, University of Melbourne, Parkville, VIC, Australia.
Histochem Cell Biol. 1997 Jan;107(1):19-29. doi: 10.1007/s004180050085.
The presence of NADPH diaphorase staining was compared with the immunohistochemical localization of four NADPH-dependent enzymes-neuronal (type I), inducible (type II), and endothelial (type III) nitric oxide synthase (NOS) and cytochrome P450 reductase. Cell types that were immunoreactive for the NADPH-dependent enzymes were also stained for NADPH diaphorase, suggesting that endothelial and neuronal NOS and cytochrome P450 reductase all show NADPH diaphorase activity in formaldehyde-fixed tissue. However, in some tissues, the presence of NADPH diaphorase staining did not coincide with the presence of any of the NADPH-dependent enzymes we examined. In vascular endothelial cells, the punctate pattern of staining observed with NADPH diaphorase histochemistry was identical to that seen following immunohistochemistry using antibodies to endothelial NOS. In enteric and pancreatic neurons and in skeletal muscle, the presence of NADPH diaphorase staining correlated with the presence of neuronal NOS. In the liver, sebaceous glands of the skin, ciliated epithelium, and a subpopulation of the cells in the subserosal glands of the trachea, zona glomerulosa of the adrenal cortex, and epithelial cells of the lacrimal and salivary glands, the presence of NADPH diaphorase staining coincided with the presence of cytochrome P450 reductase immunoreactivity. In epithelial cells of the renal tubules and zona fasciculata and zona reticularis of the adrenal cortex, NADPH diaphorase staining was observed that did not coincide with the presence of any of the enzymes. Inducible NOS was not observed in any tissue. Thus, while tissues that demonstrate immunoreactivity for neuronal and endothelial NOS also stain positively for NADPH diaphorase activity, the presence of NADPH diaphorase staining does not reliably or specifically indicate the presence of one or more NOS isoforms.
将NADPH黄递酶染色的存在情况与四种NADPH依赖性酶（神经元型一氧化氮合酶（NOS）（I型）、诱导型一氧化氮合酶（II型）、内皮型一氧化氮合酶（III型）和细胞色素P450还原酶）的免疫组织化学定位进行了比较。对NADPH依赖性酶呈免疫反应性的细胞类型也被NADPH黄递酶染色，这表明在内甲醛固定组织中，内皮型和神经元型一氧化氮合酶以及细胞色素P450还原酶均显示出NADPH黄递酶活性。然而，在某些组织中，NADPH黄递酶染色的存在情况与我们检测的任何一种NADPH依赖性酶的存在情况并不一致。在血管内皮细胞中，NADPH黄递酶组织化学观察到的点状染色模式与使用抗内皮型一氧化氮合酶抗体进行免疫组织化学后观察到的模式相同。在肠和胰腺神经元以及骨骼肌中，NADPH黄递酶染色的存在与神经元型一氧化氮合酶的存在相关。在肝脏、皮肤皮脂腺、纤毛上皮以及气管浆膜下腺、肾上腺皮质球状带、泪腺和唾液腺上皮细胞亚群中，NADPH黄递酶染色的存在与细胞色素P450还原酶免疫反应性的存在一致。在肾小管上皮细胞以及肾上腺皮质束状带和网状带中，观察到NADPH黄递酶染色，但与任何一种酶的存在情况均不一致。在任何组织中均未观察到诱导型一氧化氮合酶。因此，虽然对神经元型和内皮型一氧化氮合酶呈免疫反应性的组织也对NADPH黄递酶活性呈阳性染色，但NADPH黄递酶染色的存在并不能可靠或特异性地表明一种或多种一氧化氮合酶同工型的存在。
