Barrier disruption in the major cerebral arteries following experimental subarachnoid hemorrhage

The effects of experimental subarachnoid hemorrhage (SAH) on the blood-arterial wall barrier in the major cerebral arteries were studied in 20 normotensive dogs. Horseradish peroxidase (HRP) was given intravenously before the animals were sacrificed to assess the integrity of the barrier. Transient elevation of intracranial pressure (ICP) produced by cisternal injection of saline solution resulted in HRP leakage at the branching points of the major cerebral arteries. Extensive disturbance of the blood-arterial wall barrier was consistently observed in the major cerebral arteries after SAH, with or without elevation of ICP. These results suggest that both subarachnoid clot and a sudden rise in the ICP are important factors causing the breakdown of the blood-arterial wall barrier, but that the effect of the clot is the most profound. Electron microscopy revealed that opening of the interendothelial junctions is one of the important mechanisms responsible for the HRP leakage in the major cerebral arteries following SAH. Disturbance of arterial permeability in the major cerebral arteries following SAH probably accounts for the abnormal post-contrast enhancement that occurs in patients who are prone to develop vasospasm following aneurysm rupture, and is probably involved in the pathogenesis of vasospasm.     

基于MR血管成像比较两种兔蛛网膜下腔出血后脑血管痉挛模型

目的应用MRA对比观察一次注血法和二次注血法制造兔蛛网膜下腔出血后脑血管痉挛模型的差异。方法新西兰兔40只,随机分为对照组（5只）、一次注血组（17只）和二次注血组（18只）,分别以生理盐水（对照组）及自体动脉血（注血组）注入枕大池。于造模前和造模后1、3、5、7、9、11天行神经功能评分,采用MRA技术观测基底动脉管径的变化情况。结果 3组动物死亡率差异无统计学意义（均P〉0.05）。对照组无明显神经功能损伤,基底动脉管径造模前后差异无统计学意义（均P〉0.05）。一次注血组造模后3天神经功能损伤最明显,基底动脉于造模后1天发生收缩痉挛,于造模后3天痉挛达到高峰,管径为造模前的68.18%,后逐渐恢复正常。二次注血组造模后5天神经功能损伤症状最明显,基底动脉于造模后1天发生收缩痉挛,于造模后5天痉挛达到高峰,管径为造模前的56.72%,后逐渐恢复正常。结论兔枕大池二次注血法是制造蛛网膜下腔出血后脑血管痉挛模型的良好方法;采用MRA技术可活体观测脑血管痉挛。     

Morphologic changes in cerebral arteries after subarachnoid hemorrhage

The premise of this article is that morphologic changes observed in cerebral arteries after subarachnoid hemorrhage play an important role in the pathogenesis of associated ischemic deficits observed in this disorder. Secondly, the arteriopathic response of cerebral arteries to subarachnoid blood is similar in many respects to that observed in systemic vessels under various pathologic conditions, and common pathogenic mechanisms may exist. The data supporting these premises may be summarized as follows: 1. Morphologic changes in human and animal cerebral arteries after subarachnoid hemorrhage are temporally associated with angiographic and clinically significant vasospasm. 2. Profound morphologic changes in cerebral arteries after subarachnoid hemorrhage do not contribute to structural narrowing of the lumen through increases in vessel wall mass. Nevertheless, structural changes may act in concert with contractile mechanisms to alter normal physiologic responses and maintain a narrowed lumen. 3. The agent responsible for arterial narrowing and morphologic changes in cerebral arteries after subarachnoid hemorrhage is contained in the erythrocyte component of whole blood and is most likely hemoglobin. 4. The volume and duration of exposure of subarachnoid blood to the artery appears to be significant in the development of the angiopathic response. 5. Ultrastructural abnormalities in systemic vessels associated with hypertension, atherogenesis, and endothelial damage are similar in many respects to those seen after subarachnoid hemorrhage.     

Musical murmurs in human cerebral arteries after subarachnoid hemorrhage

A transcranial ultrasonic method for the recording of murmurs from cerebral vessels is described. Using the new approach the authors have observed musical murmurs of pure tone quality in 15 patients with increased flow velocities in the cerebral arteries after spontaneous subarachnoid hemorrhage (SAH). The frequency range of the pure tones was from 140 to 820 Hz, corresponding to flow velocities between 73 and 215 cm/sec. The musical murmurs occurred as a transitional state between silent flow and the well known phenomenon of bruit. They were observed between the 4th and the 20th day after SAH. The most likely cause of the musical murmur is a periodic shedding of vortices in the cerebral arteries, commonly referred to as "a von Kármán vortex street." Clinically the presence of musical murmurs indicated that pathologically increased blood velocities were present in the artery under investigation. This probably reflected the degree of spasm.     

Barrier disruption in the major cerebral arteries during the acute stage after experimental subarachnoid hemorrhage

The effects of experimental subarachnoid hemorrhage (SAH) on the blood-arterial wall barrier in the basilar arteries were studied during the acute stage after SAH. SAH was induced in rats by injecting fresh autologous blood into the cisterna magna. Horseradish peroxidase (HRP) was given intravenously before killing the animals to assess the integrity of the barrier. In the basilar arteries taken from the animals that were killed 30 minutes after the cisternal injection of either mock cerebrospinal fluid or arterial blood, HRP reaction products were diffusely observed in the subendothelial spaces and smooth muscle layers. At 5 hours after the blood injection, no permeation of HRP into the subendothelial space was observed. Endothelial cell transcytosis seemed to be the important mechanism for HRP permeation into the subendothelial space rather than the opening of interendothelial junctions. The disruption of the blood-arterial wall barrier in the major cerebral arteries after SAH may be involved in the pathogenesis of vasospasm.     

The significance of morphological changes in cerebral arteries after subarachnoid hemorrhage

A porcine model was developed to allow quantitative assessment of morphological changes in cerebral arteries after subarachnoid hemorrhage and to determine the significance of structural changes in producing arterial narrowing. Whole blood was selectively applied to the middle cerebral artery (MCA) of seven pigs. After 10 days, vessels were perfusion-fixed and examined by light and transmission electron microscopy and immunohistochemistry. The MCA's exposed to whole blood for 10 days showed prominent luminal narrowing associated with profound ultrastructural changes affecting all layers of the vessel wall. Morphometric analysis, however, demonstrated that significant reductions in the luminal cross-sectional area (-55.8% +/- 12.5%, p less than 0.005) and increases in radial wall thickness (75.1% +/- 10.5%, p less than 0.005) were associated with only minimal increase in the cross-sectional area of the vessel wall (12.5% +/- 15%, p less than 0.025). By stereological analysis, the volume density of individual components of the arterial wall was unchanged in MCA's exposed to blood. Vessels exposed to blood showed a 44% reduction in smooth-muscle cell immunoreactive actin and increased collagen in the extracellular matrix of the vessel wall. These data suggest that structural changes in cerebral arteries after subarachnoid hemorrhage do not directly contribute to vessel narrowing through increases in wall mass. Nevertheless, such changes may reflect pathological mechanisms which act to augment prolonged vasoconstriction or inhibit the maintenance of normal vascular tone.     

Barrier disruption in the major cerebral arteries after experimental subarachnoid hemorrhage in spontaneously hypertensive and normotensive rats

The effects of experimental subarachnoid hemorrhage (SAH) on the blood-arterial wall barrier in the major cerebral arteries were studied in 24 spontaneously hypertensive rats (SHR) and 13 Sprague-Dawley rats (SDR). Horseradish peroxidase (HRP) was given intravenously before killing the animals to assess the integrity of the barrier. In the acute experimental group, transient elevation of intracranial pressure (ICP) and systemic arterial pressure produced by cisternal injection of whole blood, saline solution, or Elliott's B solution resulted in extensive disturbance of the blood-arterial wall barrier. In the chronic group, only the cisternal injection of whole blood in SHR brought about an extensive and marked disturbance of the arterial permeability. These results suggest that: (a) early breakdown of the blood-arterial wall barrier seems to be due to a sudden rise in the ICP or arterial pressure; (b) in the chronic experiments, the subarachnoid clot is the most important factor responsible for the permeability changes; and (c) in the chronic SAH experiments, the blood-arterial wall barrier seems to be more vulnerable in SHR than in Sprague-Dawley rats. Due to the well-known similarities between SHRs and hypertensive human beings, patients with chronic hypertension should be considered at high risk after SAH for extensive blood-arterial wall barrier disturbances.     

Suramin-induced reversal of chronic cerebral vasospasm in experimental subarachnoid hemorrhage

OBJECT: The naphthylsulfonate derivative suramin is an inhibitor of growth factor receptors (receptor tyrosine kinases) and G protein-coupled P2Y receptors. Both types of these receptors are suspected of being involved in cerebral vasospasm after subarachnoid hemorrhage (SAH). In the current study, the authors examined the therapeutic effects of suramin and a selective P2X-receptor antagonist, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), in the reversal of vasospasm in an established canine double-hemorrhage model. METHODS: Twenty-four dogs underwent double blood injection into the cisterna magna, with injections given on Days 0 and 2. The dogs were divided randomly into three groups (six animals in each group) to be treated from Days 2 through 6 with the vehicle dimethyl sulfoxide, suramin, or PPADS. An additional group of six dogs received double blood injection without any treatment and served as an SAH control group. The animals were killed on Day 7. Angiography was performed on Day 0 before blood injection and again on Day 7 before the animals were killed. After the death of the animals, the basilar arteries (BAs) were collected for morphological studies and determination of tyrosine kinase expression, and the bloody cerebrospinal fluid (CSF) produced by the hemorrhages was collected for measurement of oxyhemoglobin and adenosine triphosphate (ATP). In the SAH control group, the mean diameter of the BAs on Day 7 was 46.23 +/- 6.32% of the value on Day 0 (which served as a reference of 100%). In the DMSO-treated group, the mean residual diameter of the BA was 47.77 +/- 0.8% on Day 7 compared with the value on Day 0. Suramin, but not PPADS, increased the residual diameter to 74.02 +/- 4.24% on Day 7. On Day 7 the level of ATP in the CSF was decreased and the level of oxyhemoglobin was increased, compared with values measured on Day 0. Suramin, but not PPADS, reduced tyrosine phosphorylation in the spastic BAs. CONCLUSIONS: By reducing tyrosine kinase activity, suramin may be useful in the treatment of cerebral vasospasm.     

Pharmacological studies on relaxation of spastic primate cerebral arteries in subarachnoid hemorrhage

Chronic cerebral vasospasm was induced in 16 monkeys by direct placement of a clot of autologous blood over the arteries of the circle of Willis on the right side. The middle cerebral arteries (MCA's) on the clot side all showed angiographic vasospasm, which was maximal 7 days after subarachnoid hemorrhage. Animals were sacrificed at this time and vascular responses to acetylcholine (ACh), histamine, and the calcium ionophore A23187 were studied in MCA rings from the clot (spastic) side and the non-clot (control) side. In control preparations with an intact endothelium, which had been precontracted by prostaglandin F2 alpha (PGF2 alpha), histamine and A23187 produced significant relaxation. The same concentrations of histamine and A23187 did not relax vascular tissues in which the endothelium had been mechanically removed. Acetylcholine did not produce a significant endothelium-dependent relaxation of primate MCA rings, but did relax rings of primate common carotid artery. Pretreatment with chlorpheniramine (an H1-receptor antagonist) prevented histamine-induced relaxation; however, cimetidine (an H2-receptor antagonist) had no inhibitory action. It thus seems that histamine mediates relaxation of intact MCA's mostly by an H1-receptor-mediated release of endothelium-derived relaxing factor (EDRF). Relaxations induced by histamine and A23187 in MCA's from the clot side were substantially reduced. Moreover, the small component of ACh-induced relaxation was also abolished. Endothelium-independent relaxation induced by glyceryl trinitrate (GTN) occurred in arteries from both the control and the clot sides. Constrictions induced by KC1 and PGF2 alpha were reduced on the clot side of the MCA's. These results suggest that subarachnoid hemorrhage influences both the generation of EDRF and the constriction of affected arteries. The small contraction which was elicited in spastic arteries was fully relaxed by GTN.     

Up-regulation of parathyroid hormone receptor in cerebral arteries after subarachnoid hemorrhage in monkeys

OBJECTIVE: Complementary deoxyribonucleic acid array analysis was used to determine whether vasospasm after subarachnoid hemorrhage (SAH) is associated with changes in gene expression. METHODS: Right SAHs were created in three monkeys, and the right and left middle cerebral arteries were collected 3, 7, or 14 days after SAH. Vasospasm was assessed by angiography performed on Day 0 and at tissue harvest. A complementary deoxyribonucleic acid array containing 5184 genes was used to screen for changes in gene expression by comparing the right and left middle cerebral arteries. RESULTS: There was significant expression (greater than fivefold expression of messenger ribonucleic acid compared with internal standard control) of 537 genes (10%) in the middle cerebral arteries. One hundred sixty-four genes (31%) did not change significantly, and 373 (69%) were differentially expressed at 3, 7, or 14 days after SAH. These 373 genes changed from 1.2- to 7-fold as compared with control arteries. The most common pattern was a progressive increase with increased time after SAH. The functions of differentially expressed genes included the regulation of gene expression, cell proliferation, inflammation, membrane proteins and receptors, kinases, and phosphatases. There was a marked increase in parathyroid hormone and parathyroid hormone receptor with time after SAH. Immunoblotting demonstrated a significant increase in parathyroid hormone receptor protein. CONCLUSION: The up-regulation of these proteins involved in vascular relaxation suggests that they may play a role in vasospasm. The progressive increase in messenger ribonucleic acids involved in the functions noted suggests that the pathogenesis of cerebral vasospasm involves cell proliferation, inflammation, and possibly smooth muscle phenotype change.     

Reduced platelet function in subarachnoid hemorrhage

The hypothesis that abnormalities of platelet function may relate to the occurrence or recurrence of subarachnoid hemorrhage (SAH) has been examined. Seventy patients with SAH and 65 control individuals were studied. The adenosine diphosphate (ADP) threshold for secondary platelet aggregation was significantly higher in the SAH group than in the controls. In tests using 4.0 micrograms/ml ADP, the percent platelet aggregation (at 2 minutes) and the maximum rate of platelet aggregation (over 20 seconds) were significantly lower in the SAH patients. There was no difference in total platelet count between the two groups. Platelet adhesiveness was lower in the SAH patients when compared to controls. Circulating microaggregates did not differ between the two groups. The results indicate that reduced platelet function does relate to SAH and may either contribute to aneurysmal rupture in cases of SAH or be a consequence of it.     

Effect of raloxifene on cerebral vasospasm following experimental subarachnoid hemorrhage in rats

The effect of raloxifene on cerebral vasospasm following experimental subarachnoid hemorrhage (SAH) was investigated in a rat model. Seven groups of seven rats underwent no SAH, no treatment; SAH only; SAH plus vehicle; SAH plus 3 days intraperitoneal raloxifene treatment; SAH plus 4 days intraperitoneal raloxifene treatment; SAH plus 3 days intrathecal raloxifene treatment; and SAH plus 4 days intrathecal raloxifene treatment. The basilar artery cross-sectional areas were measured at 72 or 96 hours following SAH. The results showed raloxifene decreased SAH-induced cerebral vasospasm in all treatment groups, and suggested no difference between intraperitoneal and intrathecal application, or between 3 days and 4 days of raloxifene treatment. The present study demonstrates that raloxifene is a potential therapeutic agent against cerebral vasospasm after SAH.     

Gene expression and molecular changes in cerebral arteries following subarachnoid hemorrhage in the rat

OBJECT: The authors investigated early changes in the cerebral arteries of rats that occur after subarachnoid hemorrhage (SAH). METHODS: Messenger RNA was investigated by performing microarray and quantitative real-time polymerase chain reaction (PCR) analyses, and protein expression was shown by performing immunohistochemical studies. The array data indicated that the initial processes that occur after SAH involve activation of genes involved in angiogenesis, inflammation, and extracellular matrix (ECM) remodeling. The real-time PCR investigation confirmed upregulation of genes that were observed using the microarray to be regulated, including iNOS, MMP13, and cxcl2. The authors also verified the upregulation of previously implicated genes for G-protein-coupled receptors (endothelin B [ETB], angiotensin 1 [AT1], and AT2) and metalloproteinase 9. The results of an immunohistochemical study confirmed that receptor genes that were seen to be regulated produced an increase in protein expression. Double immunostaining of rat cerebral arteries with endothelial cell- or smooth-muscle cell-specific antibodies verified that an increase in ETB, 5-hydrotryptamine (5-HT1B), and 5-HT1D receptor expression occurs in smooth-muscle cells. CONCLUSIONS: Processes occurring after SAH lead to enhanced arterial contractility and ECM remodeling either directly or through angiogenesis and inflammation. These processes are active via an increase in metalloproteinase expression, the presence of proangiogenic factors, and the expression of proinflammatory genes.     

Effects of raloxifene on cerebral vasospasm after experimental subarachnoid hemorrhage in rabbits

Background

The aim of this study was to investigate the ability of a SERM, RLX, to prevent vasospasm in a rabbit model of SAH.

Methods

Thirty-four New Zealand white rabbits were allocated into 3 groups randomly. Subarachnoid hemorrhage was induced by injecting autologous blood into the cisterna magna. The treatment groups were as follows: (1) sham operated (no SAH [n = 12]), (2) SAH only (n = 12), and (3) SAH plus RLX (n = 10). Basilar artery lumen areas and arterial wall thickness were measured to assess vasospams in all groups.

Results

There was a statistically significant difference between the mean basilar artery cross-sectional areas and the mean arterial wall thickness measurements of the control and SAH-only groups (P < .05). The difference between the mean basilar artery cross-sectional areas and the mean arterial wall thickness measurements in the RLX-treated group was statistically significant (P < .05). The difference between the SAH group and the SAH + RLX group was also statistically significant (P < .05).

Conclusions

These findings demonstrate that RLX has marked vasodilatatory effect in an experimental model of SAH in rabbits. This observation may have clinical implications suggesting that this SERM drug could be used as possible anti-vasospastic agent in patients without major adverse effects.