Extracellular Superoxide Dismutase in Cultured Astrocytes: Decrease in Cell-Surface Activity and Increase in Medium Activity by Lipopolysaccharide-Stimulation

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• 作者：Ichiro Iitsuka (1)
  Akiko Motoyoshi-Yamashiro (1)
  Mitsuaki Moriyama (1)
  Yukiko Kannan-Hayashi (1)
  Yuka Fujimoto (1)
  Katsura Takano (1)
  Koji Murakami (2)
  Yukio Yoneda (2)
  Yoichi Nakamura (1) yoichi@vet.osakafu-u.ac.jp
• 关键词：Extracellular superoxide dismutase &#8211 ; Lipopolysaccharide &#8211 ; Astrocytes &#8211 ; Cell ; surface activity of SOD &#8211 ; Neuron and microglia
• 刊名：Neurochemical Research
• 出版年：2012
• 出版时间：October 2012
• 年：2012
• 卷：37
• 期：10
• 页码：2108-2116
• 全文大小：573.1 KB
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• 作者单位：1. Laboratory of Integrative Physiology in Veterinary Science, Osaka Prefecture University, Rinku-ourai Kita 1-58, Izumisano, Osaka 598-8531, Japan2. Laboratory of Molecular Pharmacology, Division of Pharmaceutical Sciences, Kanazawa University Graduate School of Natural Science and Technology, Kanazawa, Ishikawa, Japan
• ISSN：1573-6903

文摘

Under pathological conditions such as ischemia/reperfusion, a large amount of superoxide anion (O₂ −) is produced and released in brain. Among three isozymes of superoxide dismutase (SOD), extracellular (EC)-SOD, known to be excreted outside cells and bound to extracellular matrix, should play a role to detoxify O₂ − in extracellular space; however, a little is known about EC-SOD in brain. In order to evaluate the SOD activity in extracellular space of CNS as direct as possible, we attempted to measure the cell-surface SOD activity on primary cultured rat brain cells by the inhibition of color development of a water-soluble tetrazolium due to O₂ − generation by xanthine oxidase/hypoxanthine added into extracellular medium of intact cells. The cell-surface SOD activity on cultured neuron and microglia was below the detection limit; however, that on cultured astrocyte was high enough to measure. By means of RT-PCR, all mRNA of three isozymes of SOD could be detected in the three types of the cells examined; however, the semi-quantitative analysis revealed that the level of EC-SOD mRNA in astrocytes was significantly higher than that in neurons and microglia. When astrocytes were stimulated with lipopolysaccharide (LPS) for 12–24 h, the cell-surface SOD activity decreased to a half, whereas the activity recovered after 36–48 h. The decrease in the activity was dependent on the LPS concentration. On the other hand, the SOD activity in the medium increased by the LPS-stimulation in a dose dependent manner; suggesting that the SOD protein localized on cell-surface, probably EC-SOD, was released into the medium. These results suggest that EC-SOD of astrocyte play a role for detoxification of extracellular O₂ − and the regulation of EC-SOD in astrocytes may contribute to the defensive mechanism against oxidative stress in brain.