Takenaka K, Yamada H, Sakai N, Ando T, Nakashima T, Nishimura Y
Department of Neurosurgery, Gifu University School of Medicine, Japan.
J Neurosurg. 1991 Sep;75(3):452-7. doi: 10.3171/jns.1991.75.3.0452.
The purpose of this study was to determine the effects of cerebrospinal fluid (CSF) from patients with subarachnoid hemorrhage (SAH) on cytosolic free calcium in cultured rat vascular smooth-muscle cells using the fluorescent intracellular calcium indicator fura-2/AM. Samples of CSF were collected from 12 patients (seven with and five without vasospasm) on Days 2, 6, 11, and 16 after SAH. Control CSF samples were obtained from five patients 6 to 9 months after they had undergone successful aneurysm surgery following an SAH. All CSF samples in both the non-vasospasm and vasospasm groups, regardless of the day of sampling after the SAH, induced significantly higher transient intracellular calcium elevations when compared to levels induced by control CSF. Furthermore, the addition of 2 mM ethyleneglycol-bis (beta-aminoethylether)-N,N'-tetra-acetic acid (EGTA) caused a slight reduction in the peak height in the CSF-induced intracellular calcium rise which declined more rapidly to basal levels than those studied without EGTA. In the non-vasospasm group, the intracellular calcium concentration remained stable after SAH throughout the study period. In contrast, in the vasospasm group, this concentration was highest on Day 2 post-SAH, but sharply decreased on Day 6 and rose again on Day 11. This result correlated with the clinical signs of vasospasm in these patients. These findings indicated that the intracellular calcium elevations induced by CSF obtained after SAH were due to the combination of the influx of extracellular calcium and the mobilization of intracellular calcium from storage sites. The changes in intracellular calcium concentrations in vascular smooth-muscle cells induced by CSF obtained from patients on successive days following SAH suggest that the substances that induce this repeat calcium elevation on Day 11 post-SAH may be the key spasmogens for vasospasm after SAH.
本研究的目的是使用荧光细胞内钙指示剂fura-2/AM，确定蛛网膜下腔出血（SAH）患者的脑脊液（CSF）对培养的大鼠血管平滑肌细胞胞质游离钙的影响。在SAH后的第2、6、11和16天，从12例患者（7例有血管痉挛，5例无血管痉挛）采集CSF样本。对照CSF样本取自5例在SAH后成功进行动脉瘤手术后6至9个月的患者。与对照CSF诱导的水平相比，非血管痉挛组和血管痉挛组的所有CSF样本，无论SAH后采样的日期如何，均诱导出明显更高的细胞内钙瞬态升高。此外，添加2 mM乙二醇双（β-氨基乙基醚）-N,N'-四乙酸（EGTA）导致CSF诱导的细胞内钙升高的峰值高度略有降低，且比未添加EGTA的研究更快地降至基础水平。在非血管痉挛组中，SAH后整个研究期间细胞内钙浓度保持稳定。相比之下，在血管痉挛组中，该浓度在SAH后第2天最高，但在第6天急剧下降，并在第11天再次上升。这一结果与这些患者血管痉挛的临床体征相关。这些发现表明，SAH后获得的CSF诱导的细胞内钙升高是由于细胞外钙内流和细胞内钙从储存部位动员的共同作用。SAH后连续几天从患者获得的CSF诱导的血管平滑肌细胞内钙浓度变化表明，SAH后第11天诱导这种钙重复升高的物质可能是SAH后血管痉挛的关键痉挛原。
