title: Na-K-Cl cotransporter-1 in the mechanism of cell swelling in cultured astrocytes after fluid percussion injury.
abstract: Brain edema and associated increased intracranial pressure are major consequences of traumatic brain injury (TBI). An important early component of the edema associated with TBI is astrocyte swelling (cytotoxic edema). Mechanisms for such swelling, however, are poorly understood. Ion channels/transporters/exchangers play a major role in cell volume regulation, and a disturbance in one or more of these systems may result in cell swelling. To examine potential mechanisms in TBI-mediated brain edema, we employed a fluid percussion model of in vitro barotrauma and examined the role of the ion transporter Na(+)-K(+)-2Cl(-)-cotransporter 1 (NKCC1) in trauma-induced astrocyte swelling as this transporter has been strongly implicated in the mechanism of cell swelling in various neurological conditions. Cultures exposed to trauma (3, 4, 5 atm pressure) caused a significant increase in NKCC1 activity (21%, 42%, 110%, respectively) at 3 h. At 5 atm pressure, trauma significantly increased NKCC1 activity at 1 h and it remained increased for up to 3 h. Trauma also increased the phosphorylation (activation) of NKCC1 at 1 and 3 h. Inhibition of MAPKs and oxidative/nitrosative stress diminished the trauma-induced NKCC1 phosphorylation as well as its activity. Bumetanide, an inhibitor of NKCC1, significantly reduced the trauma-induced astrocyte swelling (61%). Silencing NKCC1 with siRNA led to a reduction in trauma-induced NKCC1 activity as well as in cell swelling. These findings demonstrate the critical involvement of NKCC1 in the astrocyte swelling following in vitro trauma, and suggest that blocking NKCC1 activity may represent a useful therapeutic strategy for the cytotoxic brain edema associated with the early phase of TBI.
authors.affiliation: Department of Pathology, University of Miami School of Medicine, Miami, FL 33101, USA.
authors.forename: Arumugam R
authors.initials: AR
authors.lastname: Jayakumar
authors.forename: Kiran S
authors.initials: KS
authors.lastname: Panickar
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Curtis
authors.forename: Xiao Y
authors.initials: XY
authors.lastname: Tong
authors.forename: Mitsuaki
authors.initials: M
authors.lastname: Moriyama
authors.forename: Michael D
authors.initials: MD
authors.lastname: Norenberg
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D001929:Brain Edema / Q000209:etiology / Q000473:pathology*; D001930:Brain Injuries / Q000150:complications / Q000473:pathology; D002478:Cells, Cultured; D003497:Cyclic N-Oxides / Q000494:pharmacology; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000502:physiology; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D010474:Percussion / Q000009:adverse effects; D012333:RNA, Messenger; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D027981:Symporters / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D000096922:K Cl- Cotransporters
chemical_list: D003497:Cyclic N-Oxides; D004791:Enzyme Inhibitors; D019331:NG-Nitroarginine Methyl Ester; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D027981:Symporters; D000096922:K Cl- Cotransporters; C029217:phenyl-N-tert-butylnitrone