Cytosolic Ca2+ oscillations in human cerebrovascular endothelial cells after subarachnoid hemorrhage.
Molecular mechanisms of cerebral vasospasm after subarachnoid hemorrhage (SAH) include specific modes of cell signaling like activation of nuclear factor (NF)-kappaB and vascular cell adhesion molecules (VCAM)-1 expression. The study's hypothesis is that cisternal cerebral spinal fluid (CSF) from patients after SAH may cause Ca(2+) oscillations which induce these modes of vascular inflammation in an in vitro model of human cerebral endothelial cells (HCECs). HCECs were incubated with cisternal CSF from 10 SAH patients with confirmed cerebral vasospasm. The CSF was collected on days 5 and 6 after hemorrhage. Cytosolic Ca(2+) concentrations and cell contraction as an indicator of endothelial barrier function were examined by fura-2 microflurometry. Activation of NF-kappaB and VCAM-1 expression were measured by immunocytochemistry. Incubation of HCEC with SAH-CSF provoked cytosolic Ca(2+) oscillations (0.31+/-0.09 per min), cell contraction, NF-kappaB activation, and VCAM-1 expression, whereas exposure to native CSF had no significant effect. When endoplasmic reticulum (ER) Ca(2+)-ATPase and ER inositol trisphosphate (IP3)-sensitive Ca(2+) channels were blocked by thapsigargin or xestospongin, the frequency of the Ca(2+) oscillations was reduced significantly. In analogy to the reduction of Ca(2+) oscillation frequency, the blockers impaired HCEC contraction, NF-kappaB activation, and VCAM-1 expression. Cisternal SAH-CSF induces cytosolic Ca(2+) oscillations in HCEC that results in cellular constriction, NF-kappaB activation, and VCAM-1 expression. The Ca(2+) oscillations depend on the function of ER Ca(2+)-ATPase and IP3-sensitive Ca(2+) channels.
Scharbrodt
Wolfram
W
Department of Neurosurgery, University Hospital Giessen-Marburg, Giessen, Germany.
Abdallah
Yaser
Y
Kasseckert
Sascha A
SA
Gligorievski
Dragan
D
Piper
Hans M
HM
Böker
Dieter-K
DK
Deinsberger
Wolfgang
W
Oertel
Matthias F
MF
D000328:Adult; D000368:Aged; D015220:Calcium Channels / Q000378:metabolism; D020013:Calcium Signaling; D002478:Cells, Cultured; D018925:Chemokines / Q000378:metabolism; D019743:Chemokines, CXC; D003600:Cytosol / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism*; D005260:Female; D006706:Homeostasis; D006801:Humans; D008297:Male; D008875:Middle Aged; D016328:NF-kappa B / Q000378:metabolism; D013345:Subarachnoid Hemorrhage / Q000378:metabolism*
C508524:CXCL17 protein, human; D015220:Calcium Channels; D018925:Chemokines; D019743:Chemokines, CXC; D016328:NF-kappa B