Inhibition of calcium/calmodulin-dependent protein kinase kinase (CaMKK) exacerbates impairment of endothelial cell and blood-brain barrier after stroke.
Animals
Benzimidazoles
/ pharmacology
Blood-Brain Barrier
/ enzymology
Brain Ischemia
/ enzymology
Calcium-Calmodulin-Dependent Protein Kinase Kinase
/ antagonists & inhibitors
Cell Death
Cells, Cultured
Endothelial Cells
/ enzymology
Humans
Male
Mice, Inbred C57BL
Naphthalimides
/ pharmacology
Sirtuin 1
/ metabolism
Stroke
/ enzymology
blood-brain barrier
calcium/calmodulin-dependent protein kinase kinase
endothelial cells
sirtuin 1
stroke
Journal
The European journal of neuroscience
ISSN: 1460-9568
Titre abrégé: Eur J Neurosci
Pays: France
ID NLM: 8918110
Informations de publication
Date de publication:
01 2019
01 2019
Historique:
received:
30
03
2018
revised:
21
09
2018
accepted:
15
10
2018
pubmed:
14
11
2018
medline:
27
5
2020
entrez:
14
11
2018
Statut:
ppublish
Résumé
Brain microvascular endothelial cells play an essential role in maintaining blood-brain barrier (BBB) integrity, and disruption of the BBB aggravates the ischemic injury. CaMKK (α and β) is a major kinase activated by elevated intracellular calcium. Previously, we demonstrated that inhibition of CaMKK exacerbated outcomes, conversely, overexpression reduced brain injury after stroke in mice. Interestingly, CaMKK has been shown to activate a key endothelial protector, sirtuin 1 (SIRT1). We hypothesized that CaMKK protects brain endothelial cells via SIRT1 activation after stroke. In this study, Oxygen-Glucose Deprivation (OGD) was performed in human brain microvascular endothelial cells. Stroke was induced by middle cerebral artery occlusion (MCAO) in male mice. Knockdown of CaMKK β using siRNA increased cell death following OGD. Inhibition of CaMKK β by STO-609 significantly and selectively down-regulated levels of phosphorylated SIRT1 after OGD. Changes in the downstream targets of SIRT1 were observed following STO-609 treatment. The effect of STO-609 on cell viability after OGD was absent, when SIRT1 was concurrently inhibited. We also demonstrated that STO-609 increased endothelial expression of the pro-inflammatory proteins ICAM-1 and VCAM-1 and inhibition of CaMKK exacerbated OGD-induced leukocyte-endothelial adhesion. Finally, intracerebroventricular injection of STO-609 exacerbated endothelial apoptosis and reduced BBB integrity after 24-hr reperfusion following MCAO in vivo. Collectively, these results demonstrated that CaMKK inhibition reduced endothelial cell viability, exacerbated inflammatory responses and aggravated BBB impairment after ischemia. CaMKK activation may attenuate ischemic brain injury via protection of the microvascular system and a reduction in the infiltration of pro-inflammatory factors.
Identifiants
pubmed: 30422362
doi: 10.1111/ejn.14223
pmc: PMC6590081
mid: NIHMS1030369
doi:
Substances chimiques
Benzimidazoles
0
Naphthalimides
0
STO 609
0
Calcium-Calmodulin-Dependent Protein Kinase Kinase
EC 2.7.11.17
SIRT1 protein, human
EC 3.5.1.-
Sirtuin 1
EC 3.5.1.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
27-39Subventions
Organisme : NINDS NIH HHS
ID : R01 NS078446
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS099628
Pays : United States
Informations de copyright
© 2018 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.
Références
Proc Natl Acad Sci U S A. 2007 Sep 11;104(37):14855-60
pubmed: 17785417
Neuropharmacology. 2008 Sep;55(3):363-89
pubmed: 18308347
Stroke. 2008 Apr;39(4):1121-6
pubmed: 18323498
Cell Cycle. 2008 Jul 15;7(14):2117-22
pubmed: 18641460
Neurobiol Dis. 2008 Nov;32(2):200-19
pubmed: 18790057
J Neurosci. 2010 May 5;30(18):6398-408
pubmed: 20445066
FEBS J. 2010 Sep;277(18):3622-36
pubmed: 20659161
J Neurosci Methods. 2011 Feb 15;195(2):206-10
pubmed: 21168441
Brain. 2013 Apr;136(Pt 4):1274-87
pubmed: 23408111
Int J Mol Sci. 2013 May 24;14(6):11096-112
pubmed: 23708107
Stroke. 2013 Aug;44(8):2333-7
pubmed: 23723308
Proc Natl Acad Sci U S A. 2013 Jun 25;110(26):E2420-7
pubmed: 23754392
J Pharm Bioallied Sci. 2013 Apr;5(2):141-7
pubmed: 23833520
Stroke. 2013 Sep;44(9):2559-66
pubmed: 23868268
Cerebrovasc Dis. 2014;37(3):171-80
pubmed: 24503888
Exp Neurol. 2014 Nov;261:404-11
pubmed: 24842488
J Neurosci. 2014 Sep 3;34(36):11897-912
pubmed: 25186738
Stroke. 2014 Nov;45(11):3403-11
pubmed: 25213338
Semin Cell Dev Biol. 2015 Feb;38:35-42
pubmed: 25444848
Br J Pharmacol. 2015 Nov;172(22):5390-402
pubmed: 26362823
Front Neurosci. 2015 Oct 29;9:385
pubmed: 26578854
Neurobiol Dis. 2016 Jul;91:182-93
pubmed: 26969532
Front Cell Neurosci. 2016 Mar 04;10:56
pubmed: 26973468
Mol Brain. 2016 Jun 07;9(1):63
pubmed: 27267759
Eur J Neurosci. 2016 Aug;44(4):2139-46
pubmed: 27305894
Cancer Res. 1989 Aug 15;49(16):4435-40
pubmed: 2743332
Redox Biol. 2018 Apr;14:229-236
pubmed: 28965081
Biochim Biophys Acta Mol Basis Dis. 2018 Feb;1864(2):542-553
pubmed: 29175057