Scharbrodt Wolfram, Abdallah Yaser, Kasseckert Sascha A, Gligorievski Dragan, Piper Hans M, Böker Dieter-K, Deinsberger Wolfgang, Oertel Matthias F
Department of Neurosurgery, University Hospital Giessen-Marburg, Giessen, Germany.
J Cereb Blood Flow Metab. 2009 Jan;29(1):57-65. doi: 10.1038/jcbfm.2008.87. Epub 2008 Aug 13.
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.
蛛网膜下腔出血（SAH）后脑血管痉挛的分子机制包括特定的细胞信号传导模式，如核因子（NF）-κB的激活和血管细胞粘附分子（VCAM）-1的表达。该研究的假设是，SAH患者的脑池脑脊液（CSF）可能会引起Ca（2+）振荡，从而在人脑血管内皮细胞（HCEC）的体外模型中诱导这些血管炎症模式。将HCEC与10例确诊为脑血管痉挛的SAH患者的脑池CSF一起孵育。CSF在出血后第5天和第6天收集。通过fura-2微量荧光测定法检测胞质Ca（2+）浓度和作为内皮屏障功能指标的细胞收缩。通过免疫细胞化学测量NF-κB的激活和VCAM-1的表达。用SAH-CSF孵育HCEC会引起胞质Ca（2+）振荡（每分钟0.31±0.09次）、细胞收缩、NF-κB激活和VCAM-1表达，而暴露于天然CSF则没有显著影响。当内质网（ER）Ca（2+）-ATP酶和ER三磷酸肌醇（IP3）敏感的Ca（2+）通道被毒胡萝卜素或海绵诱捕素阻断时，Ca（2+）振荡的频率显著降低。与Ca（2+）振荡频率降低类似，这些阻滞剂损害了HCEC收缩、NF-κB激活和VCAM-1表达。脑池SAH-CSF在HCEC中诱导胞质Ca（2+）振荡，导致细胞收缩、NF-κB激活和VCAM-1表达。Ca（2+）振荡取决于ER Ca（2+）-ATP酶和IP3敏感的Ca（2+）通道的功能。
