Fasudil reduces GFAP expression after hypoxic injury

Varun Kesherwani, Shikha Tarang, Robert Barnes, Sandeep K. Agrawal

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Fasudil (HA-1077), a specific Rho kinase II (ROCKII) inhibitor, is in clinical trials for recovery from spinal cord injury (SCI). The primary role of Fasudil is in axonal regeneration, as it inhibits ROCKII, the key signaling molecule involved in collapse of axon growth cone. Astrogliosis, due to the activation of astrocytes is an indicator of CNS injury. In early stages of injury, GFAP expression increases, helping to restore the integrity of the CNS. An increase in GFAP expression is also a marker of astrogliosis. Thus, reducing GFAP and hence astrogliosis at later stages of SCI is important for neuroregeneration and functional recovery. CoCl2 was used to induce hypoxic injury in astrocytic cell lines A172 (24h) and in spinal cord dorsal column white matter (8h). Several different techniques were used to study the changes in GFAP expression such as real-time PCR, western blotting and immunofluorescence staining with confocal microscopy. Hypoxia increased the expression of GFAP in A172 cells and in the spinal cord dorsal column after CoCl2 (100μM) treatment for 24h and 8h, respectively. We observed 11 folds increase in protein expression in A172 cells (24h) and 4.5 folds in spinal cord dorsal column (8h). The RNA expression was increased 3 folds in A172 cells after 24h of treatment and 4 folds in spinal cord dorsal column after 8h of treatment with 100μM CoCl2. Treatment with fasudil (20μM) significantly reduces the expression of GFAP in A172 cells and in spinal cord dorsal column. Fasudil also decreased activation of NF-κB in A172 cells after hypoxic injury. In the present study, we observed that fasudil reduces the expression of GFAP (consequently, astrogliosis) after hypoxic injury to A172 cells and spinal cord dorsal column. Our studies demonstrate that fasudil also plays a role in GFAP expression by reducing NF-κB activation at the injury site which could further help in axonal regeneration.

Original languageEnglish (US)
Pages (from-to)45-50
Number of pages6
JournalNeuroscience Letters
Volume576
DOIs
StatePublished - Jul 25 2014

Fingerprint

Spinal Cord
Wounds and Injuries
Spinal Cord Injuries
Regeneration
rho-Associated Kinases
Growth Cones
Therapeutics
fasudil
Confocal Microscopy
Astrocytes
Fluorescent Antibody Technique
Axons
Real-Time Polymerase Chain Reaction
Western Blotting
Clinical Trials
RNA
Staining and Labeling
Cell Line
Proteins

Keywords

  • Astrogliosis
  • Fasudil
  • GFAP
  • Hypoxia
  • Spinal cord

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Fasudil reduces GFAP expression after hypoxic injury. / Kesherwani, Varun; Tarang, Shikha; Barnes, Robert; Agrawal, Sandeep K.

In: Neuroscience Letters, Vol. 576, 25.07.2014, p. 45-50.

Research output: Contribution to journalArticle

Kesherwani, Varun ; Tarang, Shikha ; Barnes, Robert ; Agrawal, Sandeep K. / Fasudil reduces GFAP expression after hypoxic injury. In: Neuroscience Letters. 2014 ; Vol. 576. pp. 45-50.
@article{7461c1a6b8434f4781214deb1c3833e1,
title = "Fasudil reduces GFAP expression after hypoxic injury",
abstract = "Fasudil (HA-1077), a specific Rho kinase II (ROCKII) inhibitor, is in clinical trials for recovery from spinal cord injury (SCI). The primary role of Fasudil is in axonal regeneration, as it inhibits ROCKII, the key signaling molecule involved in collapse of axon growth cone. Astrogliosis, due to the activation of astrocytes is an indicator of CNS injury. In early stages of injury, GFAP expression increases, helping to restore the integrity of the CNS. An increase in GFAP expression is also a marker of astrogliosis. Thus, reducing GFAP and hence astrogliosis at later stages of SCI is important for neuroregeneration and functional recovery. CoCl2 was used to induce hypoxic injury in astrocytic cell lines A172 (24h) and in spinal cord dorsal column white matter (8h). Several different techniques were used to study the changes in GFAP expression such as real-time PCR, western blotting and immunofluorescence staining with confocal microscopy. Hypoxia increased the expression of GFAP in A172 cells and in the spinal cord dorsal column after CoCl2 (100μM) treatment for 24h and 8h, respectively. We observed 11 folds increase in protein expression in A172 cells (24h) and 4.5 folds in spinal cord dorsal column (8h). The RNA expression was increased 3 folds in A172 cells after 24h of treatment and 4 folds in spinal cord dorsal column after 8h of treatment with 100μM CoCl2. Treatment with fasudil (20μM) significantly reduces the expression of GFAP in A172 cells and in spinal cord dorsal column. Fasudil also decreased activation of NF-κB in A172 cells after hypoxic injury. In the present study, we observed that fasudil reduces the expression of GFAP (consequently, astrogliosis) after hypoxic injury to A172 cells and spinal cord dorsal column. Our studies demonstrate that fasudil also plays a role in GFAP expression by reducing NF-κB activation at the injury site which could further help in axonal regeneration.",
keywords = "Astrogliosis, Fasudil, GFAP, Hypoxia, Spinal cord",
author = "Varun Kesherwani and Shikha Tarang and Robert Barnes and Agrawal, {Sandeep K.}",
year = "2014",
month = "7",
day = "25",
doi = "10.1016/j.neulet.2014.05.053",
language = "English (US)",
volume = "576",
pages = "45--50",
journal = "Neuroscience Letters",
issn = "0304-3940",
publisher = "Elsevier Ireland Ltd",

}

TY - JOUR

T1 - Fasudil reduces GFAP expression after hypoxic injury

AU - Kesherwani, Varun

AU - Tarang, Shikha

AU - Barnes, Robert

AU - Agrawal, Sandeep K.

PY - 2014/7/25

Y1 - 2014/7/25

N2 - Fasudil (HA-1077), a specific Rho kinase II (ROCKII) inhibitor, is in clinical trials for recovery from spinal cord injury (SCI). The primary role of Fasudil is in axonal regeneration, as it inhibits ROCKII, the key signaling molecule involved in collapse of axon growth cone. Astrogliosis, due to the activation of astrocytes is an indicator of CNS injury. In early stages of injury, GFAP expression increases, helping to restore the integrity of the CNS. An increase in GFAP expression is also a marker of astrogliosis. Thus, reducing GFAP and hence astrogliosis at later stages of SCI is important for neuroregeneration and functional recovery. CoCl2 was used to induce hypoxic injury in astrocytic cell lines A172 (24h) and in spinal cord dorsal column white matter (8h). Several different techniques were used to study the changes in GFAP expression such as real-time PCR, western blotting and immunofluorescence staining with confocal microscopy. Hypoxia increased the expression of GFAP in A172 cells and in the spinal cord dorsal column after CoCl2 (100μM) treatment for 24h and 8h, respectively. We observed 11 folds increase in protein expression in A172 cells (24h) and 4.5 folds in spinal cord dorsal column (8h). The RNA expression was increased 3 folds in A172 cells after 24h of treatment and 4 folds in spinal cord dorsal column after 8h of treatment with 100μM CoCl2. Treatment with fasudil (20μM) significantly reduces the expression of GFAP in A172 cells and in spinal cord dorsal column. Fasudil also decreased activation of NF-κB in A172 cells after hypoxic injury. In the present study, we observed that fasudil reduces the expression of GFAP (consequently, astrogliosis) after hypoxic injury to A172 cells and spinal cord dorsal column. Our studies demonstrate that fasudil also plays a role in GFAP expression by reducing NF-κB activation at the injury site which could further help in axonal regeneration.

AB - Fasudil (HA-1077), a specific Rho kinase II (ROCKII) inhibitor, is in clinical trials for recovery from spinal cord injury (SCI). The primary role of Fasudil is in axonal regeneration, as it inhibits ROCKII, the key signaling molecule involved in collapse of axon growth cone. Astrogliosis, due to the activation of astrocytes is an indicator of CNS injury. In early stages of injury, GFAP expression increases, helping to restore the integrity of the CNS. An increase in GFAP expression is also a marker of astrogliosis. Thus, reducing GFAP and hence astrogliosis at later stages of SCI is important for neuroregeneration and functional recovery. CoCl2 was used to induce hypoxic injury in astrocytic cell lines A172 (24h) and in spinal cord dorsal column white matter (8h). Several different techniques were used to study the changes in GFAP expression such as real-time PCR, western blotting and immunofluorescence staining with confocal microscopy. Hypoxia increased the expression of GFAP in A172 cells and in the spinal cord dorsal column after CoCl2 (100μM) treatment for 24h and 8h, respectively. We observed 11 folds increase in protein expression in A172 cells (24h) and 4.5 folds in spinal cord dorsal column (8h). The RNA expression was increased 3 folds in A172 cells after 24h of treatment and 4 folds in spinal cord dorsal column after 8h of treatment with 100μM CoCl2. Treatment with fasudil (20μM) significantly reduces the expression of GFAP in A172 cells and in spinal cord dorsal column. Fasudil also decreased activation of NF-κB in A172 cells after hypoxic injury. In the present study, we observed that fasudil reduces the expression of GFAP (consequently, astrogliosis) after hypoxic injury to A172 cells and spinal cord dorsal column. Our studies demonstrate that fasudil also plays a role in GFAP expression by reducing NF-κB activation at the injury site which could further help in axonal regeneration.

KW - Astrogliosis

KW - Fasudil

KW - GFAP

KW - Hypoxia

KW - Spinal cord

UR - http://www.scopus.com/inward/record.url?scp=84902590588&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84902590588&partnerID=8YFLogxK

U2 - 10.1016/j.neulet.2014.05.053

DO - 10.1016/j.neulet.2014.05.053

M3 - Article

C2 - 24905175

AN - SCOPUS:84902590588

VL - 576

SP - 45

EP - 50

JO - Neuroscience Letters

JF - Neuroscience Letters

SN - 0304-3940

ER -