Muse K E, Oberley T D, Sempf J M, Oberley L W
Pathology and Laboratory Medicine Service, William S. Middleton Memorial Veterans Hospital, Madison, WI 53705.
Histochem J. 1994 Sep;26(9):734-53. doi: 10.1007/BF00158205.
Immunoperoxidase and immunogold techniques were used to localize the following antioxidant enzyme systems in the adult hamster kidney at the light and ultrastructural levels: superoxide dismutases, catalases, peroxidases and glutathione S-transferases. Each cell type in the kidney showed specific patterns of labelling of these enzymes. For example, proximal and distal tubular and transitional epithelial cells showed significant staining for all of these enzymes, while glomerular cells and cells of the thin loop of Henle did not show significant staining at the light microscope level. In addition, high levels of glutathione peroxidase were found in smooth muscle cells of renal arteries. At the ultrastructural level, each enzyme was found in a specific subcellular location. Manganese superoxide dismutase was found in mitochondria, catalase was localized in peroxisomes, while copper, zinc superoxide dismutase and glutathione S-transferase (liver and placental forms) were found in both the nucleus and cytoplasm. Glutathione peroxidase was found to have a broad intracellular distribution, with localization in mitochondria, peroxisomes, nucleus, and cytoplasm. Microvilli of tubular cells were labelled by antibodies to catalase, copper, zinc superoxide dismutase, glutathione peroxidase, and glutathione S-transferases. Cell types that were negative by light microscopy immunoperoxidase studies showed definite labelling with immunogold post-embedding ultrastructural techniques (glomerular cells and cells of the loop of Henle), demonstrating the greater sensitivity of the latter technique. These observations demonstrate that there are large variations in the levels of antioxidant enzymes in different cell types, and that even within a distinct cell type, the levels of these enzymes vary in different subcellular locations. Our results demonstrate for the first time the overall antioxidant enzyme status of individual kidney cell types, thereby explaining why different cell types have differing susceptibilities to oxidant stress. Possible physiological and pathological consequences of these findings are discussed.
超氧化物歧化酶、过氧化氢酶、过氧化物酶和谷胱甘肽S-转移酶。肾脏中的每种细胞类型都显示出这些酶的特定标记模式。例如,近端和远端肾小管及移行上皮细胞对所有这些酶都有明显染色,而肾小球细胞和髓袢细段细胞在光镜水平未显示明显染色。此外,在肾动脉平滑肌细胞中发现了高水平的谷胱甘肽过氧化物酶。在超微结构水平上,每种酶都位于特定的亚细胞位置。锰超氧化物歧化酶存在于线粒体中,过氧化氢酶定位于过氧化物酶体,而铜锌超氧化物歧化酶和谷胱甘肽S-转移酶(肝脏和胎盘形式)存在于细胞核和细胞质中。谷胱甘肽过氧化物酶在细胞内分布广泛,定位于线粒体、过氧化物酶体、细胞核和细胞质中。肾小管细胞的微绒毛被抗过氧化氢酶、铜锌超氧化物歧化酶、谷胱甘肽过氧化物酶和谷胱甘肽S-转移酶的抗体标记。光镜免疫过氧化物酶研究呈阴性的细胞类型在用免疫金包埋后超微结构技术时显示出明确的标记(肾小球细胞和髓袢细胞),表明后一种技术具有更高的敏感性。这些观察结果表明,不同细胞类型中抗氧化酶的水平存在很大差异,而且即使在同一特定细胞类型内,这些酶在不同亚细胞位置的水平也有所不同。我们的结果首次证明了单个肾细胞类型的整体抗氧化酶状态,从而解释了为什么不同细胞类型对氧化应激的敏感性不同。讨论了这些发现可能的生理和病理后果。