Browse > Article
http://dx.doi.org/10.4196/kjpp.2013.17.4.275

Expression and Activity of the Na-K ATPase in Ischemic Injury of Primary Cultured Astrocytes  

Kim, Mi Jung (Department of Physiology, Biomedical Science Institute and Medical Research Center, School of Medicine, Kyung Hee University)
Hur, Jinyoung (Department of Physiology, Biomedical Science Institute and Medical Research Center, School of Medicine, Kyung Hee University)
Ham, In-Hye (Department of Physiology, Biomedical Science Institute and Medical Research Center, School of Medicine, Kyung Hee University)
Yang, Hye Jin (Department of Physiology, Biomedical Science Institute and Medical Research Center, School of Medicine, Kyung Hee University)
Kim, Younghoon (Department of Physiology, Biomedical Science Institute and Medical Research Center, School of Medicine, Kyung Hee University)
Park, Seungjoon (Department of Pharmacology, Biomedical Science Institute and Medical Research Center, School of Medicine, Kyung Hee University)
Cho, Young-Wuk (Department of Physiology, Biomedical Science Institute and Medical Research Center, School of Medicine, Kyung Hee University)
Publication Information
The Korean Journal of Physiology and Pharmacology / v.17, no.4, 2013 , pp. 275-281 More about this Journal
Abstract
Astrocytes are reported to have critical functions in ischemic brain injury including protective effects against ischemia-induced neuronal dysfunction. Na-K ATPase maintains ionic gradients in astrocytes and is suggested as an indicator of ischemic injury in glial cells. Here, we examined the role of the Na-K ATPase in the pathologic process of ischemic injury of primary cultured astrocytes. Chemical ischemia was induced by sodium azide and glucose deprivation. Lactate dehydrogenase assays showed that the cytotoxic effect of chemical ischemia on astrocytes began to appear at 2 h of ischemia. The expression of Na-K ATPase ${\alpha}1$ subunit protein was increased at 2 h of chemical ischemia and was decreased at 6 h of ischemia, whereas the expression of ${\alpha}1$ subunit mRNA was not changed by chemical ischemia. Na-K ATPase activity was time-dependently decreased at 1, 3, and 6 h of chemical ischemia, whereas the enzyme activity was temporarily recovered to the control value at 2 h of chemical ischemia. Cytotoxicity at 2 h of chemical ischemia was significantly blocked by reoxygenation for 24 h following ischemia. Reoxygenation following chemical ischemia for 1 h significantly increased the activity of the Na-K ATPase, while reoxygenation following ischemia for 2 h slightly decreased the enzyme activity. These results suggest that the critical time for ischemia-induced cytotoxicity of astrocytes might be 2 h after the initiation of ischemic insult and that the increase in the expression and activity of the Na-K ATPase might play a protective role during ischemic injury of astrocytes.
Keywords
Chemical ischemia; Na-K ATPase; Primary cultured astrocytes;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Benarroch EE. Neuron-astrocyte interactions: partnership for normal function and disease in the central nervous system. Mayo Clin Proc. 2005;80:1326-1338.   DOI   ScienceOn
2 Swanson RA, Ying W, Kauppinen TM. Astrocyte influences on ischemic neuronal death. Curr Mol Med. 2004;4:193-205.   DOI   ScienceOn
3 Stanimirovic DB, Ball R, Durkin JP. Stimulation of glutamate uptake and Na,K-ATPase activity in rat astrocytes exposed to ischemia-like insults. Glia. 1997;19:123-134.   DOI
4 Swanson RA, Benington JH. Astrocyte glucose metabolism under normal and pathological conditions in vitro. Dev Neurosci. 1996;18:515-521.   DOI
5 Silver IA, Erecińska M. Energetic demands of the$Na^{+}$/$K^{+}$ ATPase in mammalian astrocytes. Glia. 1997;21:35-45.   DOI
6 Xie M, Wang W, Kimelberg HK, Zhou M. Oxygen and glucose deprivation-induced changes in astrocyte membrane potential and their underlying mechanisms in acute rat hippocampal slices. J Cereb Blood Flow Metab. 2008;28:456-467.   DOI   ScienceOn
7 Kwon HJ, Hwang IK, An HJ, Han SH, Yang JI, Shin HS, Yoo ID, Kang TC. Changes of glial $Na^{+}$-$K^{+}$ ATPase (alpha 1 subunit) immunoreactivity in the gerbil hippocampus after transient forebrain ischemia. Brain Res. 2003;987:233-239.   DOI   ScienceOn
8 McCarthy KD, de Vellis J. Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol. 1980;85:890-902.   DOI   ScienceOn
9 Selvatici R, Previati M, Marino S, Marani L, Falzarano S, Lanzoni I, Siniscalchi A. Sodium azide induced neuronal damage in vitro: evidence for non-apoptotic cell death. Neurochem Res. 2009;34:909-916.   DOI
10 Bhuiyan MI, Kim HB, Kim SY, Cho KO. The Neuroprotective Potential of Cyanidin-3-glucoside Fraction Extracted from Mulberry Following Oxygen-glucose Deprivation. Korean J Physiol Pharmacol. 2011;15:353-361.   DOI   ScienceOn
11 Hur J, Lee P, Kim MJ, Kim Y, Cho YW. Ischemia-activated microglia induces neuronal injury via activation of gp91phox NADPH oxidase. Biochem Biophys Res Commun. 2010;391: 1526-1530.   DOI   ScienceOn
12 Scherzer P, Popovtzer MM. Segmental localization of mRNAs encoding $Na^{+}$-$Ka^{+}$-ATPase alpha(1)-and beta(1)-subunits in diabetic rat kidneys using RT-PCR. Am J Physiol Renal Physiol. 2002;282:F492-500.
13 Mitsumoto Y, Klip A. Development regulation of the subcellular distribution and glycosylation of GLUT1 and GLUT4 glucose transporters during myogenesis of L6 muscle cells. J Biol Chem. 1992;267:4957-4962.
14 Esmann M. ATPase and phosphatase activity of $Na^{+}$, $K^{+}$- ATPase: molar and specific activity, protein determination. Methods Enzymol. 1988;156:105-115.   DOI
15 Molinas SM, Trumper L, Serra E, Elías MM. Evolution of renal function and $Na^{+}$, $K^{+}$-ATPase expression during ischaemiareperfusion injury in rat kidney. Mol Cell Biochem. 2006;287: 33-42.   DOI
16 Skou JC. The Na,K-pump. Methods Enzymol. 1988;156:1-25.   DOI
17 Di Carlo A. Human and economic burden of stroke. Age Ageing. 2009;38:4-5.
18 Dugan LL, Kim-Han JS. Astrocyte mitochondria in in vitro models of ischemia. J Bioenerg Biomembr. 2004;36:317-321.   DOI   ScienceOn
19 Rossi DJ, Brady JD, Mohr C. Astrocyte metabolism and signaling during brain ischemia. Nat Neurosci. 2007;10:1377- 1386.   DOI   ScienceOn
20 Golden WC, Brambrink AM, Traystman RJ, Martin LJ. Failure to sustain recovery of Na,K-ATPase function is a possible mechanism for striatal neurodegeneration in hypoxic-ischemic newborn piglets. Brain Res Mol Brain Res. 2001;88:94-102.   DOI   ScienceOn
21 Hernández-R J. $Na^{+}$/$K^{+}$-ATPase regulation by neurotransmitters. Neurochem Int. 1992;20:1-10.
22 Specht SC. Development and regional distribution of two molecular forms of the catalytic subunit of the $Na^{+}$, $K^{+}$ -ATPase in rat brain. Biochem Biophys Res Commun. 1984;121:208-212.   DOI   ScienceOn
23 Kasai K, Yamashita T, Yamaguchi A, Yoshiya K, Kawakita A, Tanaka H, Sugimoto H, Tohyama M. Induction of mRNAs and proteins for Na/K ATPase alpha1 and beta1 subunits following hypoxia/reoxygenation in astrocytes. Brain Res Mol Brain Res. 2003;110:38-44.   DOI   ScienceOn
24 Watts AG, Sanchez-Watts G, Emanuel JR, Levenson R. Cellspecific expression of mRNAs encoding $Na^{+}$,$K^{+}$-ATPase alpha-and beta-subunit isoforms within the rat central nervous system. Proc Natl Acad Sci U S A. 1991;88:7425-7429.   DOI   ScienceOn
25 Bezzi P, Volterra A. A neuron-glia signalling network in the active brain. Curr Opin Neurobiol. 2001;11:387-394.   DOI   ScienceOn