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Rho-Kinase Inhibitor, Fasudil, Prevents Neuronal Apoptosis via the Akt Activation and PTEN Inactivation in the Ischemic Penumbra of Rat Brain

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Abstract

Recently, some studies suggested that inhibition of Rho-kinase (ROCK) prevented cerebral ischemia injury through inhibiting inflammatory reaction, increasing cerebral blood flow, modulating the neuronal actin cytoskeleton polymerization, and preventing tau hyperphosphorylation and p25/CDK5 increase. However, there is little information regarding the effects of ROCK inhibitor on the neuronal apoptosis in ischemic brain injury. In this study, we determined whether ROCK inhibitor, fasudil, inhibited ischemic neuronal apoptosis through phosphatase and tensin homolog deleted on chromosome10 (PTEN)/Akt/signal pathway in vivo. Adult male Sprague–Dawley rats were subjected to permanent middle cerebral artery occlusion. Rats received ROCK inhibitor, fasudil (10 mg/kg), at 30 min before middle cerebral artery occlusion. The infarct area, neuronal apoptosis and caspase-3 activity was significantly decreased by fasudil with improvement of neurological deterioration. However, the beneficial effects of fasudil were attenuated by the co-application of LY294002 (PI3K inhibitor). Fasudil maintained postischemic Akt activity at relatively proper level and decreased the augmentation of PTEN and ROCK activity in the penumbra area. Furthermore, fasudil inhibited attenuation of GSK-β and Bad phosphorylation in the penumbra area. In conclusion, the findings provide another consideration that fasudil protects the brain against ischemia injury through decreasing neuronal apoptosis and reveals the link between the ROCK inhibition and the PTEN/Akt pathway.

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Abbreviations

Bad:

Bcl-2 associated death protein

CBF:

Cerebral blood flow

HRP:

Horseradish peroxidase

MCAO:

Middle cerebral artery occlusion

MYPT:

Myosin-binding subunit of myosin light chain phosphatase

PI3K:

Phosphatidylinositol 3 kinase

PIP3:

Phosphatidylinositol (PI)-3,4,5-triphosphate

GSK-3:

Glycogen synthase kinase-3

PTEN:

Phosphatase and tensin homolog deleted on chromosome 10

RGCs:

Retinal ganglion cells

ROCK:

Rho-kinase

TBST:

Tris-buffered saline Tween-20

TTC:

2,3,5-Triphenyltetrazolium chloride

TUNEL:

Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling

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Acknowledgments

This work was supported by the National Nature Science Foundation of China [30672457 to Y. X.F. and 30971428 to D.F.W.].

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Authors and Affiliations

  1. Department of Pharmacy, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China

    Jianhua Wu, Hankun Hu, Ping Liu & Dongfang Wu

  2. Department of Pharmacology, School of Pharmaceutical Sciences, Central South University, Changsha, 410078, China

    Yunxiang Fang

  3. Department of Pharmacy, Ruikang Hospital, Guangxi College of Traditional Chinese Medicine, Nanning, 530011, Guangxi, China

    Jianzhe Li

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  1. Jianhua Wu
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Correspondence to Dongfang Wu.

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Wu, J., Li, J., Hu, H. et al. Rho-Kinase Inhibitor, Fasudil, Prevents Neuronal Apoptosis via the Akt Activation and PTEN Inactivation in the Ischemic Penumbra of Rat Brain. Cell Mol Neurobiol 32, 1187–1197 (2012). https://doi.org/10.1007/s10571-012-9845-z

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