Hydrocephalus after intraventricular hemorrhage: the role of thrombin

Previous studies demonstrated that thrombin is an important factor in brain injury after intracerebral hemorrhage. This study investigated the effect of thrombin on hydrocephalus development in a rat intraventricular hemorrhage (IVH) model. There were three parts in this study. First, male Sprague–Dawley rats had an injection of 200 μL saline, autologous blood or heparinized blood, into the right lateral ventricle. Second, rats had an injection of 50 μL saline or 3U thrombin into the right lateral ventricle. Third, rats had an injection of thrombin (3U) with a protease-activated receptor-1 (PAR-1) antagonist, {"type":"entrez-protein","attrs":{"text":"SCH79797","term_id":"1052762130","term_text":"SCH79797"}}SCH79797 (0.15 nmol), or vehicle into the right lateral ventricle. Lateral ventricle volumes were measured by magnetic resonance imaging and the brains were used for immunohistochemistry and western blot analyses. Intraventricular injection of autologous blood induced hydrocephalus from day 1 to 28. Heparinized blood injection resulted in less hydrocephalus at all time points compared with blood injection alone (P<0.05). Intraventricular injection of thrombin caused significant hydrocephalus, ventricular wall damage, and periventricular blood–brain barrier disruption. Thrombin-induced hydrocephalus was reduced by co-injection of the PAR-1 antagonist {"type":"entrez-protein","attrs":{"text":"SCH79797","term_id":"1052762130","term_text":"SCH79797"}}SCH79797 (P<0.05). In conclusion, thrombin contributes to hydrocephalus development after IVH and thrombin-induced hydrocephalus is through PAR-1.     

大鼠脑室出血后脑铁代谢与脑积水发生关系的研究

目的 观察脑室出血后铁离子和铁蛋白的动态变化,及去铁敏治疗前后慢性脑积水的发生率.方法 SD雄性大鼠170只,随机分为5组:正常对照组、假性脑室出血组、脑室出血组、侧脑室注铁离子组及脑室出血加去铁敏组.以前肉后0.4 mm冠状面的侧脑室宽度评价脑积水的发生率,以亚铁嗪法测量大鼠脑脊液铁离子含量,以ELISA试剂盒测量脑组织铁蛋白含量.结果 正常对照组、假性脑室出血组无脑积水发生,脑室出血组、侧脑室注铁离子组、脑室出血加去铁敏组28 d时脑积水发生率分别为80％、73％、20％,脑室出血加去铁敏组脑积水发生率显著低于脑室出血组和侧脑室注铁离子组(P＜0.01).脑脊液中铁离子含量和脑组织中铁蛋白在脑室出血组和侧脑室注铁离子组中各个时相点均显著高于假性脑室出血组(P＜0.01);脑室出血加去铁敏组铁离子含量在各个时相点,及铁蛋白在7、28d时均显著低于脑室出血组(P＜0.01).结论 大鼠脑室出血后脑脊液和脑组织铁含量明显升高,去铁敏可降低脑脊液中铁离子和脑组织中铁蛋白含量,降低脑积水的发生率.     

Intraventricular fibrinolysis with tissue plasminogen activator is associated with transient cerebrospinal fluid inflammation: a randomized controlled trial

Locally administered tissue plasminogen activator (TPA) accelerates clearance of intraventricular hemorrhage (IVH), but its impact on neurologic outcomes remains unclear and preclinical research suggests it may have pro-inflammatory effects. We randomly allocated patients with ruptured cerebral aneurysms and IVH, treated with endovascular coiling and ventricular drainage, to receive either 2-mg intraventricular TPA or placebo every 12 hours. Cerebrospinal fluid (CSF) and serum cytokine and white blood cell (WBC) concentrations were measured before drug administration and daily for 72 hours. Cerebrospinal fluid D-dimer levels were assessed 6 and 12 hours after administration to quantify fibrinolysis. Six patients were randomized to each group. Patients treated with TPA developed higher CSF cytokine concentrations compared with placebo-treated patients (P<0.05 for tumor necrosis factor-α, interferon-γ, interleukin (IL)-1α, IL-1β, IL-2, IL-4, and IL-6), as well as higher CSF WBC counts (P=0.03). Differences were greatest after 24 hours and decreased over 48 to 72 hours. The magnitude of the inflammatory response was significantly associated with peak CSF D-dimer concentration and extent of IVH clearance. We conclude that intraventricular TPA administration produces a transient local inflammatory response, the severity of which is strongly associated with the degree of fibrinolysis, suggesting it may be induced by release of hematoma breakdown products, rather than the drug itself.     

去铁敏对大鼠脑室出血后慢性脑积水与脑组织Wnt1、Wnt3a表达的影响

目的 观察去铁敏对大鼠脑室出血(IVH)后脑积水的干预效果及脑组织Wnt1、Wnt3a的表达影响.方法 雄性SD大鼠130只随机分为正常组、假性IVH组、IVH组及IVH加去铁敏组.采用脑室出血后慢性脑积水大鼠模型,于术后1 d、7 d及28 d处死大鼠,观察脑积水的发生率和脑组织中Wnt基因及蛋白表达情况.结果 正常组、假性IVH组未见脑积水发生,IVH组28 d脑积水发生率80%(12/15),显著高于IVH加去铁敏组28 d脑积水发生率20%(3/15).IVH后Wnt1、Wnt3amRNA及蛋白表达在7 d及28 d均显著高于假性IVH组,而去铁敏治疗组Wnt1、Wnt3a mRNA及蛋白表达在28 d较IVH组均显著降低.结论 去铁敏治疗可降低大鼠IVH后慢性脑积水发生率,Wnt信号通路参与了IVH后脑积水发生及去铁敏干预机制.

Abstract：

Objective To observe the effect of deferoxamine on chronic hydrocephalus after intraventricular hemorrhage (IVH) and the role of Wnt (Wnt1 and Wnt3a). Methods A total of 130Sprague Dawley male rats were randomly assigned into 4 groups: normal control group, sham - IVH group,IVH group and deferoxamine - treated group. The rats of subgroups except normal control group received an injection of autologous blood or saline into right lateral cerebral ventricles. Deferoxamine or vehicle was administered 3 hours after IVH and then every 12 hours up to 7 days in IVH group and deferoxaminetreated group. Rats were euthanized at 1, 7, 28 days for measurement, and the transverse diameter of the lateral ventricles were observed for evaluation of hydrocephalus. The expression of Wnt1 and Wnt3a were measured by RT -PCR and Western Blot. Results At 28 days, there was no hydrocephalus in the normal control group and sham - IVH group. 80 %(12/15) of the rats in IVH group developed hydrocephalus. The rate was only 20 % ( 3/15 ) in deferoxamine - treated group, which was much lower than that in IVH group. The expression of Wnt1 mRNA was normal in normal control group and shame - IVH group, and upregulated in IVH group. The peak expression was detected at 28 days. The expression was significantly higher than that in shame - IVH group at 7 days and 28 days, after deferoxamine treatment, downregulation of Wnt1 mRNA were observed and were significantly lower than that in IVH group. The Wnt3a mRNA had similar trends, while the expressed level was normal in normal control group and shame - IVH group, and upregulated following IVH. The peak expression was detected at 28 days, and was significantly higher than that in shame - IVH group at 7 days and 28 days. After deferoxamine treatment, the downregulation of Wnt3a mRNA was observed and was significantly lower than that in IVH group. Accordingly, Wnt1 protein expression was normal in normal control group and shame - IVH group, and increased after IVH. At 7 days and 28 days, the Wnt1 protein expression of IVH group was markedly higher than shame - IVH group. After deferoxamine treatment, the expression of Wnt1 protein was reduced and significantly lower than that of IVH group. The expression of Wnt3a protein was also normal in normal control group and shame - IVH group. The up-regulation of the protein expression was observed following IVH at 28 days, while down - regulation was observed after deferoxamine treatment at 28 days. Conclusions Deferoxamine could reduce hydrocephalus after IVH, and Wnt pathway may play an important role in the development of IVH and therapeutic effect of deferoxamine.     

去铁敏对大鼠脑室出血后慢性脑积水与脑组织Wnt1、Wnt3a表达的影响

目的 观察去铁敏对大鼠脑室出血(IVH)后脑积水的干预效果及脑组织Wnt1、Wnt3a的表达影响.方法 雄性SD大鼠130只随机分为正常组、假性IVH组、IVH组及IVH加去铁敏组.采用脑室出血后慢性脑积水大鼠模型,于术后1 d、7 d及28 d处死大鼠,观察脑积水的发生率和脑组织中Wnt基因及蛋白表达情况.结果 正常组、假性IVH组未见脑积水发生,IVH组28 d脑积水发生率80%(12/15),显著高于IVH加去铁敏组28 d脑积水发生率20%(3/15).IVH后Wnt1、Wnt3amRNA及蛋白表达在7 d及28 d均显著高于假性IVH组,而去铁敏治疗组Wnt1、Wnt3a mRNA及蛋白表达在28 d较IVH组均显著降低.结论 去铁敏治疗可降低大鼠IVH后慢性脑积水发生率,Wnt信号通路参与了IVH后脑积水发生及去铁敏干预机制.     

去铁敏对大鼠脑室出血后慢性脑积水与脑组织Wnt1、Wnt3a表达的影响

目的 观察去铁敏对大鼠脑室出血(IVH)后脑积水的干预效果及脑组织Wnt1、Wnt3a的表达影响.方法 雄性SD大鼠130只随机分为正常组、假性IVH组、IVH组及IVH加去铁敏组.采用脑室出血后慢性脑积水大鼠模型,于术后1 d、7 d及28 d处死大鼠,观察脑积水的发生率和脑组织中Wnt基因及蛋白表达情况.结果 正常组、假性IVH组未见脑积水发生,IVH组28 d脑积水发生率80%(12/15),显著高于IVH加去铁敏组28 d脑积水发生率20%(3/15).IVH后Wnt1、Wnt3amRNA及蛋白表达在7 d及28 d均显著高于假性IVH组,而去铁敏治疗组Wnt1、Wnt3a mRNA及蛋白表达在28 d较IVH组均显著降低.结论 去铁敏治疗可降低大鼠IVH后慢性脑积水发生率,Wnt信号通路参与了IVH后脑积水发生及去铁敏干预机制.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

大鼠脑室系统出血模型的建立

目的 探讨建立稳定、制作过程简单、创伤小的大鼠脑室系统出血模型的方法 . 方法 取大鼠自体动脉血立体定向下注入右侧侧脑室建立大鼠腩室系统出血模型,对模型组和对照组大鼠在不同时间点进行神经行为学评分,并观察脑室及室周脑组织病理变化. 结果本方法 模型成功率为88.9%(16/18),注血后6h大鼠出现行为异常,7d后行为异常好转;光镜下观察发现注血后24h模型组大鼠室管膜连续性遭到破坏,细胞间隙增宽,室周脑组织轻度水肿及出现红色坏死神经元. 结论 本研究采用的造模方法 模型稳定,制作创伤小,病理变化接近临床.     

Edaravone Reduces Iron-Mediated Hydrocephalus and Behavioral Disorder in Rat by Activating the Nrf2/HO-1 Pathway

Our previous studies have demonstrated that hemorrhage-derived iron has a key role in causing brain injury after intraventricular hemorrhage (IVH). Based on this finding, we hypothesized that edaravone, a free-radical scavenger, has the potential to alleviate hydrocephalus and neurological deficits post-IVH by suppressing iron-induced oxidative stress. Thus, this study aimed to investigate the efficacy of edaravone for rats with FeCl₃ injection, as well as to explore the related molecular mechanism. An experimental model was established in adult male Sprague-Dawley rats via FeCl₃ injection into the right lateral ventricle. Edaravone or vehicle was administered immediately, 1 day and 2 days after intraventricular injection. Brain water content, magnetic resonance imaging, neurological score, oxidative stress assays, Western blot analysis, and electron microscopy were employed to evaluate brain injury in these rats. Intraventricular injection of FeCl₃ induced brain edema, ventricular dilation, and neurobehavioral disorder in rats. Edaravone treatment significantly attenuated the above symptoms, reduced ependymal cilia and neuron damage, and inhibited oxidative stress (elevated levels of an antioxidant, superoxide dismutase; decreased levels of an oxidant, malondialdehyde). Moreover, edaravone administration effectively activated the Nrf2/HO-1 signaling pathway in rat brain following FeCl₃ injection. These results showed that edaravone treatment alleviated brain edema, ventricular expansion, and neurological disorder after FeCl₃ injection. The possible mechanism is by protecting ependymal cilia and neurons from oxidative stress injury and activating the Nrf2/HO-1 signaling pathway. These results provide further experimental evidence for edaravone application in the treatment of IVH.     

Tissue plasminogen activator for the treatment of intraventricular hematoma: the dose-effect relationship

In this study, we investigated the dose-effect relationship and safety of tissue plasminogen activator (tPA) for the treatment of intraventricular hemorrhage/hematoma (IVH) in rats. Adult male Sprague-Dawley rats were injected with autologous blood into the left lateral ventricle to establish IVH. Two hours later, Ringer's saline or 0.25-2 microg of tPA were administered directly to the IVH over 3 h. The regional cerebral blood flow (rCBF) on the surface of the left parietal cortex was measured with laser Doppler flowmetry. Twenty-four hours after the build-up of IVH, the brains were removed for morphometrical and histological studies. A dose of 0.5-2 microg tPA significantly diminished the IVH in a dose-dependent manner (p < 0.001). However, only the dose of 0.5 microg tPA significantly ameliorated the reduction of rCBF 24 h after IVH (p < 0.01). TPA did not improve the ventricular dilatation on the side with IVH. Instead, 1-2 microg of tPA caused additional injuries, including intraventricular leukocytosis and edema of periventricular tissues and choroid plexus on both hemispheres. These results indicate that higher doses of tPA may have detrimental effects on the brain. The dosage rate of 0.5 microg seems beneficial to treat 5 microl of IVH (equals to a dose of 0.1 mg/ml blood) in our model in terms of the satisfactory fibrinolysis and less damage to the brain.     

Iron toxicity in mice with collagenase-induced intracerebral hemorrhage

Intracerebral hemorrhage (ICH) is a devastating form of stroke. In this study, we examined the efficacy of deferoxamine (DFX), an iron chelator, after collagenase-induced ICH in 12-month-old mice. Intracerebral hemorrhage was induced by intrastriatal injection of collagenase. Deferoxamine (200 mg/kg, intraperitoneal) or vehicle was administrated 6 hours after ICH and then every 12 hours for up to 3 days. Neurologic deficits were examined on days 1 and 3 after ICH. Mice were killed after 1 or 3 days of DFX treatment for examination of iron deposition, neuronal death, oxidative stress, microglia/astrocyte activation, neutrophil infiltration, brain injury volume, and brain edema and swelling. Collagenase-induced ICH resulted in iron overload in the perihematomal region on day 3. Systemic administration of DFX decreased iron accumulation and neuronal death, attenuated production of reactive oxygen species, and reduced microglial activation and neutrophil infiltration without affecting astrocytes. Although DFX did not reduce brain injury volume, edema, or swelling, it improved neurologic function. Results of our study indicate that iron toxicity contributes to collagenase-induced hemorrhagic brain injury and that reducing iron accumulation can reduce neuronal death and modestly improve functional outcome after ICH in mice.     

术中超声引导神经内镜手术与脑室外引流术治疗脑室内血肿伴铸型的对比分析研究

目的对比分析术中超声引导神经内镜手术和脑室外引流术治疗脑室血肿伴铸型的安全性及有效性。方法选取我科2016年5月至2018年11月间脑室血肿伴铸型的患者60例,根据脑室血肿伴铸型的类型分为三型,采取分层数字随机法将患者分成两组,对照组(n=30)采取传统的脑室外引流术治疗,实验组(n=30)则采取术中超声引导神经内镜下脑室内血肿清除术,比较两组临床疗效。结果实验组术后24 h血肿清除率、术后24小时GCS评分、术后6月Barthel日常生活能力评分均明显高于对照组(P0.05),实验组引流管带管时间、住院时间较对照组缩短,同时降低死亡率(P0.05);实验组术区再出血、分流依赖性脑积水、颅内感染与对照组比较无统计学差异(P 0.05)。结论术中超声引导下神经内镜手术治疗脑室血肿伴铸型是安全、有效的,该治疗方案优于脑室外引流术。     

Morphological changes following experimental intraventricular haemorrhage and intraventricular fibrinolytic treatment with recombinant tissue plasminogen activator

Intraventricular haemorrhage (IVH) occurs in up to 50% of patients with primary intracerebral haemorrhage and aneurysmal subarachnoid haemorrhage. It is a significant and independent contributor to mortality and morbidity in these intracranial haemorrhages. Using a model of isolated IVH, we assessed the morphological changes induced by intraventricular bleeding and investigated the effects of intraventricular fibrinolytic treatment following IVH. IVH was induced in 32 pigs by intraventricular infusion of 10 ml autologous blood along with thrombin. The treatment group received an intraventricular injection of 1.5 mg (1 mg/ml) tissue plasminogen activator (tPA) following the injection of blood. The placebo group received the same volume of normal saline. Morphological examinations of the brains were carried out 7 days and 6 weeks following IVH. The ventricles were incompletely filled with blood and significantly enlarged in the placebo group 7 days after the IVH. In contrast, no residual intraventricular clots were visible in the animals treated with tPA, and the diameters of the lateral ventricles had returned to normal within 7 days. Marked losses of the ependymal covering of the ventricular walls were found in the placebo-treated animals, while the ependymal layer was largely intact in the animals treated with tPA. No haemorrhages induced by tPA were observed. The results indicate that intraventricularly administered tPA significantly enhances the lysis of intraventricular blood clots, accelerates the resolution of acute posthaemorrhagic hydrocephalus, and preserves the integrity of the ependymal layer. Received: 6 October 1999 / Revised, accepted: 26 January 2000