casein-kinase-ii and Infarction--Middle-Cerebral-Artery

Topics: Animals; beta Catenin; Casein Kinase II; Disease Models, Animal; Gene Expression Regulation; Gene Knockdown Techniques; Glycosides; GTP Phosphohydrolases; Humans; Infarction, Middle Cerebral Artery; Ischemic Stroke; Lymphoid Enhancer-Binding Factor 1; Mice; Mitochondrial Dynamics; Multiprotein Complexes; Neuroprotection; Nuclear Proteins; T Cell Transcription Factor 1; Transcription Factors; Transcription, Genetic

5d, a novel analogue of the racemic 3-n-butylphthalide (NBP), has been reported for its free radical scavenging activity in vitro and preventive neuroprotection in vivo. Nevertheless, the mechanism by which 5d attenuated ischemia/reperfusion (I/R) injury is still unknown. Our results showed that 5d significantly increased CK2 activity as well as CK2α and 2α' protein levels after I/R injury. Besides, 5d suppressed the translocation of cytosolic p47phox and Rac1 to the membrane, decreased NOX4 expression and ROS generation. Furthermore, 5d blocked the dissociation between CK2α and Rac1 so as to decrease NADPH oxidase activity. Based on these findings, we propose that the neuroprotective effect of 5d is due to an increase of CK2 activity, which blocks I/R-induced dissociation between CK2α and Rac1, decreases NADPH oxidase activity, inhibits ROS production and finally realizes the neuroprotection of I/R. These findings point to that 5d might be considered an attractive candidate for further studies in ischemic stroke.

Topics: Animals; Benzofurans; Brain; Casein Kinase II; Cell Membrane; Cell Survival; Cytosol; Gene Expression Regulation, Enzymologic; Infarction, Middle Cerebral Artery; Male; NADPH Oxidases; Neurons; Neuroprotective Agents; Oxidative Stress; rac1 GTP-Binding Protein; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Reperfusion Injury; RNA Interference

This study shows the in vivo neuroprotective effect of cilostazol against cerebral ischemic injury evoked by subjecting rats to 2-h occlusion of middle cerebral artery (MCAO) followed by 24-h reperfusion. We observed the signaling pathway by which cilostazol suppressed MCAO-induced increased phosphorylation of phosphatase and tensin homolog deleted from chromosome 10 (PTEN) and apoptosis via increased phosphorylation of casein kinase 2 (CK2). When rats received 30 mg/kg cilostazol orally two times at 5 min and 4 h after the completion of ischemia, the infarct area was significantly reduced in the cortex and striatum with improvement of neurological deterioration. Increased DNA fragmentation in the penumbral zone was significantly reduced by cilostazol. Cilostazol significantly elevated phosphorylation levels of CK2, Akt, and cyclic AMP response element-binding protein (CREB) in association with increased Bcl-2 in the ischemic area, whereas the elevated PTEN phosphorylation was significantly reduced, all of which were antagonized by iberiotoxin, a maxi-K channel blocker, administered intracisternally 30 min before ischemia. In conclusion, cilostazol ameliorates the neuronal damage by suppression of apoptotic cell death via the maxi-K channel opening-coupled up-regulation of CK2 phosphorylation and down-regulation of PTEN phosphorylation with resultant increase in the Akt and CREB phosphorylation and increased Bcl-2 protein.

Topics: Animals; Blood-Brain Barrier; Blotting, Western; Brain Ischemia; Casein Kinase II; Chromosomes; Cilostazol; Cyclic AMP Response Element-Binding Protein; DNA Fragmentation; Infarction; Infarction, Middle Cerebral Artery; Microfilament Proteins; Neuroprotective Agents; Phosphoric Monoester Hydrolases; Phosphorylation; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Tensins; Tetrazoles; Water