Pifithrin-α reduces cerebral vasospasm by attenuating apoptosis of endothelial cells in a subarachnoid haemorrhage model of rat

Background The mechanism of cerebral vasospasm following subarachnoid haemorrhage （SAH） is not understood. Here, we hypothesized that apoptosis of endothelial cells induced by p53 and its target gene em dash p53 upregulated modulator of apoptosis （PUMA） played an important role in development of cerebral vasospasm. We also observed the effects of a p53 inhibitor, pifithrin-α （PFT-α）, on reducing the expression of p53 and PUMA, consequently decreasing the apoptosis of endothelial cells and alleviating cerebral vasospasm. Methods Male Sprague-Dawley rats weighing 300-350 g were randomly divided into five groups： a control group （sham surgery）, a SAH group, a SAH＋dimethyl sulfoxide （DMSO） group, a SAH＋PFT-α （0.2 mg/kg） group and a SAH＋PFT-α （2.0 mg/kg） group. PFT-α was injected intraperitoneally immediately after SAH. Rats were sacnficed 24 hours after SAH. Western blot and immunohistochemical staining were used to detect the levels of p53, PUMA and caspase-3 protein. In addition, mortality and neurological scores were assessed for each group. Statistical significance was assured by analysis of variance performed in one way ANOVA followed by the Tukey test. The neurological and mortality scores were analyzed by Dunn＇s method and Fisher exact test, respectively. Results After SAH, Western blot and immunohistochemical staining showed the levels of p53, PUMA and caspase-3 in the endothelial cells and the numbers of TdT mediated dUTP nick end labelling （TUNEL） positive endothelial cells were all significantly increased in the basilar arteries （P 〈0.05）, but significantly reduced by PFT-a （P 〈0.05）. These changes were accompanied by increasing diameters and declining wall thickness of basilar arteries （P〈O.05）, as well as reduced mortality and neurological deficits of the rats （P〈O.05）. Conclusions PFT-a could protect cerebral vessels from development of vasospasm and improve neurological outcome as well as reduce the mortality via suppressing apoptos     

Protective Effects of Overexpression of bcl-xl Gene on Local Cerebral Infarction in Transgenic Mice Undergoing Permanent Occlusion of Middle Cerebral Artery

In order to investigate the protective effects of the overexpression of bcl-xl gene on local cerebral infarction in the transgenic mice subject to permanent occlusion of middle cerebral artery, the models of bcl-xl transgenic mice were established and subjected to cerebral infarction by intraluminal occlusion of the middle cerebral artery. The infarct volume and the neurological scores were observed and comparison between the wild type mice and the transgenic mice was made. It was found that the infarct volume and the neurological scores in the transgenic mice were significantly decreased as compared with those in the wild type mice. It was suggested that the overexpression of bcl-xl gene in transgenic mice could reduce the infarct volume and improve the neurological function of the mice.     

EFFECTS OF TRANSCRANIAL MAGNETIC STIMULATION ON MOTOR CORTICAL EXCITABILITY AND NEUROFUNCTION AFTER CEREBRAL ISCHEMIA-REPERFUSION INJURY IN RATS

Objective To clarify the effects of repetitive transcranial magnetic stimulation （rTMS） on rat motor cortical excitability and neurofunction after cerebral ischemia-reperfusion injury. Methods After determined awake resting motor threshold （MT） and motor evoked potentials （MEPs） of right hindlimbs, 20 Sprague-Dawley rats were subjected to middle cerebral artery occlusion （MCAO） reperfusion injury, then rTMS were applied to rTMS group （n = 10） at different time, while control group （n = 10） received no stimulation. A week later, MT and MEPs were evaluated again, as well as neurological deficits and infarct volume. The effects of rTMS and MCAO reperfusion injury on these parameters were analyzed. Results： After MCAO reperfusion, both MT level and neurological deficit scores increased, distinct focal infarction formed, and latency of MEP elongated. Compared with the control group, the increased extent of MT and neurological scores of rats receiving rTMS were significantly lower （P〈 0.05）, as well as the infarct volumes reduced significantly（P 〈 0.05）. But MEP was not affected by rTMS obviously. There was a positive linear correlation between postinjury MT and infarct volume （r = 0.64, P 〈 0.05）. Conclusion rTMS may facilitate neurofunction recovery after cerebral ischemia-reperfusion. Postinjury MT could provide prognostic information after MCAO reperfusion injury.     

Immune Vasculitis Induced Atherosclerosis

Summary： The relationship between immune vasculitis and atherosclerosis was studied. The experimental model of weanling rabbits for immune vasculitis was reproduced by intravenous injection of 10% bovine serum albumin. There were 6 groups： group A, 25 weanling rabbits with immune vasculitis subject to coronary arteriography; group B, 10 normal mature rabbits subject to coronary arteriography; group C, 10 weanling rabbits subject to coronary arteriography, group D, 8 weanling rabbits with vasculitis and cholesterol diet; group E, 8 weanling rabbits receiving single cholesterol diet; group F： 8 weanling rabbits receiving basic diet. Four weeks later, coronary arteriography was performed in groups A, B and C. The rabbits in groups D, E and F were sacrificed for the study of pathological changes in the coronary artery after 12 weeks. The results showed that the dilatation of coronary artery occurred in 6 rabbits of group A, but in groups B and C, no dilatation of coronary artery appeared. In comparison with group E, more severe atherosclerosis occurred in group D, showing the thickened plaque, fibrous sclerosis and atherosclerotic lesion. Percentage of plaques covering aortic intima, incidence of atherosclerosis of small coronary arteries and degree of stenosis of coronary arteries were significantly higher in group D than in group E （P〈0.01）. No atherosclerosis changes were found in group F. It was concluded that in the acute phase, the serum immune vasculitis can induce the dilatation of coronary artery of some weanling rabbits, and aggravate the formation of atherosclerosis in rabbits fed with cholesterol diet. Immune vasculitis is a new risk factor of atherosclerosis and ischemic heart disease.     

Nimodipine Modulates Bcl-2 and Bax mRNA Expression after Cerebral Ischemia

In order to explore whether the member of Bcl-2 gene family, for example, Bcl-2 and Bax, are induced after cerebral ischemia, and whether expression of genes can be modulated by calcium-antagonist, the rat cerebral ischemic models were made by occluding left middle cerebral artery. The expression of Bcl-2 and Bax mRNA was measured by RT-PCR method. After middle cerebral artery occlusion (MCAO), the expression of both Bcl-2 and Bax mRNA were induced. Level of Bcl-2 mRNA increased steadily and level of Bax mRNA increased gradually at first, reached a peak after 24 h, then decreased slowly. After administration of nimodipine, Bcl-2 mRNA was up-regulated in the hippocampus 6 and 24h after ischemia, while Bax mRNA was down-regulated 6 and 24 h after ischemia. Focal cerebral ischemia can induce proto-oncogenes to express, which was associated with apoptosis. Calcium-antagonist can up-regulate Bcl-2 mRNA and down-regulate Bax mRNA.The increased ratio of Bcl-2 and Bax mRNA may contribute to the antiapoptic effect of nimodipine.The study indicates that pharmacological modulation of Bcl-2 family member expression could become a new stratehgy to manage neuronal damage.     

Fenestrated basilar artery associated with multiple aneurysms

A 59-year-old male presented with a history of feeling somewhat "funny" and having an episode of slurred speech. No neurological deficits were found. Multislice CT and CTA demonstrated multiple aneurysms associated with fenestrated basilar artery.     

Changes of endothelin during cerebral vasospasm after experimental subarachnoid hemorrhage.

Laser-Doppler flowmetry was employed in the observation of regional cortical blood flow (rCBF) after experimental subarachnoid hemorrhage (SAH) in anesthetized rats and the contents of endothelin (ET) in the cerebral-spinal fluid (CSF), plasma, hypothalamus, cerebral cortex, cerebellum and medulla were simultaneously measured. There was a biphasic change of rCBF after SAH. The contents of ET in CSF, plasma and hypothalamus rose prominently in the early stage after SAH, and the ET-increase in CSF and hypothalamus was earlier than that in plasma. The changes of ET contents in CSF correlated well with that in hypothalamus. Our results suggest that ET probably is an early important factor which induces cerebral vasospasm (CVS) after SAH. The increase of ET in CSF, which may originate from the hypothalamus, might play a more important role in the development of CVS after SAH than that in plasma.     

MECHANISM OF PATHOLOGICAL CHANGES OF INTRAVENTRICULAR HEMORRHAGE IN DOGS

Objective To probe the mechanism of pathological changes of intraventricular hemorrhage (IVH). Methods The evaluation of neurological status, serial CT scans and pathological examination were applied on the canine model of lVH. The ventricular volume and blood clot volume were measured based on the CT images. Results The normal adult canine ventricle tend to be slitlike. After injection, the ventricle was obviously dilated by the blood clot. The linear regression of ventricular volume against blood clot volume was significant in the first week. From then on, however, while the clots continued to shrink, the ventricular volume showed progressive elargement. The clots were lysed completely within 3 to 4 weeks. The linear regression of the degree of ventricular dilatation against the first clot volume was also significant. In the pathological examination ,we found the ependyrnal lining of ventricular system was destroyed and neurons in the subependymal areas developed acidophil necrosis, which was prominent around Sylvian aqueduct. Conclusion Hemorrhagic ventricular dilatation (HVD) is a prominent feature of IVH and also is a strong indicator for poor prognosis. lschemic changes of periventricular neurons in some important structures may be the most direct cause for poor outcome of IVH. It may be induced by periventricular vascular structures compressed by HVD, increased intracranial pressure, cerebral vasospasm and others.     

NEUROSURGERY

4.1 Cerebrovascular disorder 206070 Experiment study of intrathecal infusion of pa-paverine for the prevention of vasospasm following sub-arachnoid hemorrhage/Li Jun(李俊, Dept Neurosurg, Wuhan Gener Hosp, Wuhan 430070)… // Chin J Exp Surg. -2005,22(12),-1528～1529 To evaluate the effect of intrathecal infusion of papaverine for the prevention of vasospasm following subarachnoid hemorrhage(SAH). Methods A cerebral vasospasm model was established in rabbits by injecting self artery blood into cistern. Vasospasm was diagnosed by Doppler identifying blood flow velocity(BFV)being increased more than 10 cm/s. Results After intrathecal infusion of papaverine, BFV was decreased from(23.     

脑血管痉挛后体感诱发电位潜伏期变化和尼莫地平的影响

目的 探讨尼莫地平对蛛网膜下腔出血（ＳＡＨ）后脑血管痉挛所致神经功能损伤的保护作用。方法 对单纯ＳＡＨ组和尼莫地平处理组大鼠观察手术前后基底动脉管径,并检测局部脑血流量（ｒＣＢＦ）、体感诱发电位（ＳＥＰ）及脑组织内皮素－１（ＥＴ－１）含量的动态变化。结果 ＳＡＨ组大鼠在诱导ＳＡＨ后ｒＣＢＦ立即降低,并持续２４ｈ,同时有基底动脉痉挛;ＳＡＨ后１ｈ开始至２４ｈＳＥＰ潜伏期逐渐延长,脑组织ＥＴ－１含量显     

人组织激肽释放酶对兔脑血管痉挛的影响

目的 探讨人组织激肽释放酶(HTK)对兔蛛网膜下腔出血(SAH)后症状性脑血管痉挛(CVS)的影响.方法 建立兔CVS模型,将双侧颈总动脉结扎2周后存活的兔子随机分为4组:假手术(Sham)组、蛛网膜下腔出血(SAH)组、HTK组,尼莫地平(ND)组.除Sham组外,其余3组动物行二次枕大池注血.后两组于第1次注血后第1～5天分别经静脉给HTK或ND.所有动物于第6天处死.用三维CT血管造影(3D-CTA)观察各组注血前后基底动脉直径变化,并对比基底动脉病理改变.结果 与SAH组相比,HTK组基底动脉痉挛不明显(P<0.01),光镜见基底动脉病理改变不明显,ND组基底动脉痉挛无明显缓解(P>0.05).结论 SAH后早期应用人组织激肽释放酶对兔脑血管痉挛具有明显的改善作用.     

电针配合尼莫地平对兔蛛网膜下腔出血后症状性脑血管痉挛的影响

[目的]观察电针联合尼莫地平治疗对兔蛛网膜下腔出血（SAH）后症状性脑血管痉挛（DCVS）的影响.[方法]采用Endo 法建立兔DCVS模型,40只随机分为模型组、电针组、药物组、针药组.电针组取＂百会＂、＂大椎＂、＂曲池＂、＂足三里＂穴,药物组用按0.1mg·kg-1·d-1静脉注射尼莫地平,治疗7d.观察各组动物食量、神经功能评分,用放射免疫法检测脑脊液（CSF）中ET、CGRP的含量,用化学法检测CSF中NO的含量,用经颅多普勒超声仪（TCD）检测基底动脉的血流速度.[结果]模型组兔食量明显降低,神经症状明显;而电针、药物及针药组食量减少、神经症状均较模型组程度轻（P＜0.05）,而针药组又较电针、药物组为轻（P＜0.05）.在SAH后第7天检测CSF中ET、NO、CGRP三项指标见差异性明显（P＜0.05）.观察针药组脑血管痉挛程度较电针、药物组减轻（P＜0.05）.[结论]电针和尼莫地平均能改善兔症状性脑血管痉挛的神经症状,但针药联合缓解血管痉挛的效果较单纯电针和尼莫地平更好.     

兔蛛网膜下腔出血后症状性脑血管痉挛模型的建立及继发脑损伤评估

目的建立日本大耳白兔蛛网膜下腔出血（SAH）后症状性脑血管痉挛模型。方法结扎双侧颈动脉后枕大池二次注血方法制成兔SAH后症状性脑血管痉挛模型。采用随机数字表法将兔分为SAH组和对照组,每组12只。采用三维CT血管造影（3D—CTA）检测SAH前后基底动脉直径,并观察进食量、神经功能变化,5d后取海马分别行电镜观察与超氧化物歧化酶（SOD）、丙二醛（MDA）测定。结果与对照组比较,SAH组兔出现严重的神经功能障碍,5d后基底动脉出现明显痉挛,海马神经元出现核固缩,线粒体空泡样变等神经损伤的改变,SOD活性降低,MDA含量升高。结论该模型可以作为SAH后症状性脑血管痉挛的实验模型。     

兔脑血管痉挛后海马CA1区c-FOS表达及尼莫地平的神经保护作用

目的 :研究脑血管痉挛 (cerebralvasospasm ,CVS)后兔海马CA1区神经元c FOS蛋白表达和尼莫地平 (ND)的神经保护作用 .方法 :枕大池二次注血法制备兔CVS模型 ,2 8只新西兰兔随机分为 3组 :假手术 (Sham)组 4只 ,蛛网膜下腔出血 (subarachnoidhemorrhage,SAH)组和ND治疗组各 1 2只 .数字减法血管造影术 (digitalsubtractionangiography ,DSA)测量CVS前后兔基底动脉 (BA)直径 ,于二次注射后 2h ,2 4h ,3d ,7d各时相点处死 ,HE染色观察海马CA1区的病理改变 ,免疫组织化学染色检测c FOS蛋白的表达 .结果 :与Sham组相比 ,2h ,2 4h ,3d ,7d时SAH组和ND组BA直径明显减小 (P <0 .0 5 ) ;3d ,7d时海马CA1区内正常神经元计数明显减少 (P <0 .0 5 ) ;2h ,2 4h ,3d时c FOS表达明显增多 (P <0 .0 5 ) ;与SAH组相同时点比较 ,ND组海马CA1区内正常神经元计数明显增多 (P <0 .0 5 ) ,c FOS表达明显减少 (P <0 .0 5 ) .结论 :c fos基因参与脑血管痉挛所致迟发性脑缺血 ,ND对其缺血损伤有神经保护作用 ,并能够减少c FOS蛋白表达     

兔脑血管痉挛前后基底动脉中PDEⅤ表达变化

目的：探讨兔蛛网膜下腔出血脑血管痉挛前后基底动脉中PDEⅤ的表达变化。方法：制作脑血管痉挛动物模型,空白对照组、实验对照组与SAH组均于制模后第3天行TCD与选择性椎-基底动脉造影,检测兔脑血管痉挛的发生及变化,检测完后,取其基底动脉,制成病理切片备用。通过免疫组化检测兔基底动脉中PDEⅤ的表达。结果：各配伍组中TCD测得的基底动脉血流速度、DSA测得的基底动脉管径值及免疫组化阳性表达产物的SUM值不等或不全相等（P〈0.05）,经Student-Newman-Keuls检验对各配伍组数据进行两两比较,SAH组与空白对照组、SAH组与实验对照组TCD测得的基底动脉血流速度、DSA测得的基底动脉管径值及免疫组化阳性表达产物的SUM值不等（P〈0.05）,空白对照组与实验对照组TCD测得的基底动脉血流速度、DSA测得的基底动脉管径值及免疫组化阳性表达产物的SUM值差异无统计学意义（P〉0.05）。结论：兔蛛网膜下腔出血脑血管痉挛后PDEⅤ在其基底动脉中明显表达,推测PDEⅤ表达可能与脑血管痉挛发生有关。     

兔SAH后NF-κB与脑血管内皮细胞凋亡及脑血管痉挛的关系

目的:探讨NF-κB在兔SAH后脑血管内皮细胞凋亡和脑血管痉挛的关系。方法:选用健康清洁新西兰白兔40只,采用二次枕大池注血法建立家兔SAH模型。白兔随机分为五组,分别为3 d组,7 d组,14 d组,干预组及对照组,每组8只。干预组为在7 d时间点注血后加入NF-κB的抑制剂PDTC,对照组为空白对照。实验兔在相应时间点分别处死,采用HE染色观察兔基底动脉血管腔直径的改变和管壁厚度的变化、免疫组织化学法观察兔基底动脉血管壁组织病理改变和NF-κB的表达、用末端标记法(TUNEL)分析兔基底动脉内皮细胞凋亡的变化。结果:SAH后第3天基底动脉血管腔已开始狭窄,管壁增厚,第7天达到高峰,之后渐减轻;干预组与SAH 7 d组比较血管壁变化明显缓解(P<0.05);SAH后3 d NF-κB表达增加,7 d表达最为明显,14 d表达稍减少;干预组基底动脉NF-κB表达较SAH 7 d组明显下降(P<0.05);SAH 7 d组基底动脉细胞凋亡指数较正常对照组显著升高(P<0.05);干预组基底动脉细胞凋亡指数较SAH 7 d组明显下降(P<0.05)。结论:NF-κB促进SAH后脑血管内皮细胞凋亡,NF-κB抑制剂PDTC可以通过抑制细胞凋亡缓解CVS。     

蛛网膜下腔出血后脑血管痉挛模型制作的优化与观察

目的明确蛛网膜下腔出血(SAH)模型经改良后脑血管痉挛(CVS)双相变化时程发展及与缺血损伤的关系。方法新西兰大白兔随机分为对照组和注血后不同时间点模型组。用优化的经皮一次注血法制作SAH模型,取脑组织冠状切视交叉前后2～4mm和中段基底动脉制备组织病理切片,HE染色后测定动脉皱缩指数(CC)和海马CA1区正常神经元细胞。每日行神经行为学评分,观察血凝块并评分。结果动物苏醒时间短。血管皱缩从注血后12h开始出现至第4d均明显,第5d时有所缓解后再次加重(P<0.05);海马CA1区存活神经元密度在各组间比较差异无统计学意义(P>0.05);CC和海马CA1区存活神经元数量间无相关关系。结论本改良模型短时、平稳,注血后12h直至第4d属于急性脑血管痉挛,第5d后属于迟发性脑血管痉挛,暂不适宜用海马CA1区神经细胞作SAH后脑损伤的病理评价。     

EPA与尼莫地平对蛛网膜下腔出血后脑血管痉挛影响的对比研究

目的:探讨二十碳五烯酸(EPA)与尼莫地平(nimodipine)对减轻动脉瘤性蛛网膜下腔出血后的脑血管痉挛的影响。方法:选70例接受开颅夹闭(craniotomy and clip)手术治疗的动脉瘤性蛛网膜下腔出血患者,随机分为对照组(n1=16),尼莫地平组(n2=26)和EPA组(n3=28)。对照组患者术后未特殊服用解痉药物,EPA组和尼莫地平组患者术后第4天开始口服1800mg/d剂量的EPA和尼莫地平120mg/d。观察脑血管迟发痉挛、脑梗死发生及术后一月随访的临床结果。结果:EPA和尼莫地平可以显著减少脑血管痉挛(n1group:36.1%,n2group:14.5%,P<0.05;n3group:13.8%,P<0.05)和脑血管痉挛引起的脑梗死的发生(n1group:26%,n2group:4.2%,P<0.01;n3group:3.5%,P<0.01),而EPA组和尼莫地平组1个月临床随访结果不良事件无明显差异。结论:EPA仅为减轻动脉瘤性蛛网膜下腔出血后的脑血管痉挛和脑梗死提供了一个治疗方向,但其具体机制仍需进一步研究。     

p38MAPK抑制剂对兔脑血管痉挛病理变化的影响

目的 观察兔脑血管痉挛(CVS)后基底动脉病理学改变、p38MAPK的表达及其抑制剂SB203580对脑血管痉挛的影响,以探讨蛛网膜下腔出血(SAH)后CVS细胞信号通路转导机制.方法 ①采用二次枕大池注血建立兔CVS模型.②采用免疫组织化学技术动态观察兔基底动脉血管壁组织病理改变,观察使用SB203580干预后对病理...