CCK-8对家兔SAH后迟发性脑血管痉挛病理变化的影响

目的:利用胆囊收缩素-8(CCK-8)的抗炎作用,观察其对自发性蛛网膜下腔出血后迟发性脑血管痉挛病理改变的影响。方法:雄性新西兰白兔60只随机分为四组(n=15)。①假手术组:仅进行枕大池穿刺和假注血;②SAH组:经家兔枕大池二次注射自体动脉血(0．8ml／kg)制作SAH模型;③SAH+CCK-8组:对SAH模型自day 0开始经枕大池注入 CCK-8(8μg／kg,0．5ml),每日一次直到动物处死;④SAH+生理盐水组:对SAH模型自day 0开始经枕大池注入等量37℃生理盐水,每日一次直到动物处死。各组分别在day 4、day 7和day 14分三批处死动物,每批5只。结果:假手术组动物基底动脉组织结构正常。SAH组动物基底动脉在day4、day7时出现血管痉挛,以day 7时最为显著,day 14血管痉挛得到缓解。CCK-8治疗组动物的血管痉挛程度有不同程度缓解,血管壁NF-κB、TNF-a表达明显减弱,与同时段SAH组和SAH+ 生理盐水组比较以day7时最为显著(P<0．05)。结论:CCK-8对SAH后迟发性脑血管痉挛具有一定的预防作用。     

川芎嗪对大鼠蛛网膜下腔出血后脑血管痉挛病理变化的影响

目的探讨川芎嗪对大鼠蛛网膜下腔出血(SAH)后CVS病理变化的影响。方法健康成年SD大鼠6 0只随机分为四组(n=1 5)。①A为NS组:枕大池两次穿刺注入生理盐水②B为SAH+NS组③C为SAH+N imotop组④D为SAH+TMP组。B、C、D三组均在枕大池两次注入自体动脉血制作成SAH模型后,自当天开始分别经腹腔注人生理盐水2 m l,或尼莫同0.1 mg,或川芎嗪2 0 mg,每日一次直到动物处死。各组动物分别在第1、4、7天分三批处死,每批5只。结果 A组动物基底动脉组织结构正常。B组动物基底动脉在第1天时出现血管痉挛,第4天时出现血管壁增厚,均以第7天时最为显著。C组动物血管痉挛程度有不同程度缓解,但第4天亦出现血管壁增厚,在第7天明显。D组动物血管痉挛及血管壁增厚明显缓解,血管壁NF-кBp6 5,Cox2的表达与同时间段B组和C组比较明显减弱(P<0.0 5)。结论川芎嗪对SAH脑血管痉挛具有一定的预防作用。     

兔蛛网膜下腔出血后基底动脉NF-κB、ICAM-1表达的研究

目的探讨兔蛛网膜下腔出血(SAH)后核转录因子-κB(NF-κB)及细胞间粘附分子-1(ICAM-1)在脑血管痉挛(CVS)发病机制中的作用及其相互间内在关系。方法新西兰纯种大白兔160只,枕大池二次注血,制作兔SAH后CVS模型。实验动物随机分为对照组、生理盐水(NS)组和SAH后3个亚组(3d、7d、11d组)。分离基底动脉,应用形态学观察、免疫组化和原位杂交等方法,观察基底动脉(BA)管径、血管壁上NF-κB及ICAM-1表达的动态变化。结果对照组及NS组脑血管造影BA光滑、平直,无串珠样改变;SAH后第3d,DSA显示在椎基底动脉交接处可见狭窄;第7d,BA管腔狭窄明显。伴随着血管腔管径的变化,血管壁上NF-κB及ICAM-1的表达也出现相应的变化。NF-κB与ICAM-1在蛋白质和mRNA水平上的表达强弱有时间上的先后性,在CVS发生的早期即出现NF-κB的表达升高,继而出现ICAM-1的高表达,而ICAM-1的表达强弱恰好与CVS的发生、发展过程相一致。对照组和NS组在BA的内皮细胞上仅在局部有微弱的表达。结论NF-κB、ICAM-1参与了痉挛血管壁的炎症反应,且其表达的变化可能与CVS的发生和发展有关。     

蛛网膜下腔血性与非血性刺激物对脑血管影响的组织学比较

目的通过对蛛网膜下腔血性、非血性刺激物对脑血管影响的比较,探讨蛛网膜下腔出血(SAH)后迟发性脑血管痉挛(DCVS)的发病机制。方法将40只新西兰白兔随机分为5组:假手术组、非抗凝自体动脉血组、抗凝自体动脉血组、Kaoline(高岭土)组和失活细菌组,每组8只。假手术组动物仅做枕大池假穿刺;非抗凝血组动物采用经枕大池二次注血法制作DCVS模型;抗凝自体动脉血组用肝素处理后的自体动脉血代替非抗凝血行枕大池二次注血;Kaoline组和失活细菌组经枕大池穿刺分别注入15%Kaoline悬浊液(0.25ml/kg)、失活细菌悬浊液(3×1010个/ml;0.25ml/kg)。各组受试动物在7d时用4%多聚甲醛灌注处死,将脑组织连同基底动脉及其分支取出行HE染色、免疫组化检测TNF-α供组织学研究。结果与假手术组基底动脉血管形态相比,非抗凝血组、Kaoline组和失活细菌组均有不同程度的血管痉挛,抗凝血组无明显血管痉挛。TNF-α表达随血管痉挛程度加重而增加。结论蛛网膜下腔内不同刺激物导致与SAH后DCVS相同的病理改变提示:蛛网膜下腔内刺激物所致血管局部的过度炎症反应可能是导致血管痉挛发生的重要原因。     

甲基强的松龙对兔实验性脑血管痉挛的作用

目的探讨大剂量甲基强的松龙对蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)的作用。方法将24只雄性新西兰白兔随机分成2组:SAH对照组和SAH 大剂量甲基强的松龙(MP,18mg/kg)治疗组。通过枕大池二次注血法构建SAH模型,观察MP对脑基底动脉的影响。应用酶联免疫生化技术检测各组兔基底动脉血管平滑肌细胞膜蛋白激酶C(PKC)活性。结果经脑血管造影证实该剂量甲基强的松龙明显减轻实验性脑血管痉挛的严重程度,与对照组相比,PKC活性在大剂量甲基强的松龙治疗组没有明显提高。结论大剂量甲基强的松龙能够明显减轻脑血管痉挛程度,通过抑制血管平滑肌细胞来防治脑血管痉挛的发生发展。     

eNOS与蛛网膜下腔出血后脑血管痉挛的关系

目的 探讨内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)在蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后不同时间点基底动脉表达情况及其与脑血管痉挛的关系.方法 新西兰大白兔72只,随机分成两组:(1)对照组(12只);(2)SAH组(60只).SAH组进一步按时间随机分为术后1d、3d、5d、7d、10d5个亚组,每组12只.采用枕大池二次注血法建立兔SAH模型.二次注血后在各个时间点采用灌流固定法处死动物,测定基底动脉横截面积判断有无脑血管痉挛.应用Western blotting分别观察eNOS在基底动脉中的表达情况.结果 枕大池二次注血法成功制作SAH模型,基底动脉发生了不同程度的血管痉挛.Western blotting观察到eNOS在对照组大量表达,实验组第1天表达开始减少,第5天表达水平最低,之后表达逐渐增加.结论 SAH后eNOS表达水平的变化与脑血管痉挛发展的时相性具有一定的一致性,eNOS蛋白减少可能参与蛛网膜下腔出血后脑血管痉挛的发展.     

巴曲酶鞘内注射防治犬蛛网膜下腔出血后迟发性脑血管痉挛的实验研究

目的　探讨巴曲酶鞘内注射防治蛛网膜下腔出血后迟发性脑血管痉挛 (SAH DCVS)及其所致脑水肿的作用。方法　“枕大池二次注血法”建立犬SAH DCVS动物模型 ,采用DSA等方法动态观察了巴曲酶鞘内注射对犬SAH DCVS的脑血管口径大小的影响 ,干湿法测定脑组织含水量变化。结果　巴曲酶组在枕大池注血后 7d脑血管口径显著大于单纯注血组 (P <0 .0 1 ) ,而脑组织含水量则显著低于单纯注血组 (P <0 .0 1 )。结论　巴曲酶可以防治SAH DCVS及其所致的脑水肿     

MS-CTA容积重建技术评价兔脑SAH后迟发性脑血管痉挛

目的动态评价多层螺旋CT血管成像(MS-CTA)容积重建技术(VR)在迟发性脑血管痉挛(DCVS)的应用价值。方法22只日本大耳白兔,采用枕大池二次注血法制作兔脑基底动脉DCVS模型,分别在第1、4、7、14d行MS-CTA检查后,立即处死动物并在光镜下测量基底动脉直径。MS-CTA使用GE Lightspeed pro 16层螺旋CT扫描仪,原始图像三维后处理采用容积重建(VR)技术。结果MS-CTA VR图像上,DCVS在第1d出现,第7d达到高峰,第14d可见基底动脉痉挛有一定程度缓解,光镜下基底动脉直径的时相变化与MS-CTA相似。结论MS-CTA VR能快速准确地评价DCVS的血管动态变化。     

甲基强的松龙对兔实验性脑血管痉挛的作用

目的探讨大剂量甲基强的松龙对蛛网膜下腔出血（SAH）后脑血管痉挛（CVS）的作用。方法将24只雄性新西兰白兔随机分成2组：SAH对照组和SAH＋大剂量甲基强的松龙（MP，18mg／kg）治疗组。通过枕大池二次注血法构建SAH模型，观察MP对脑基底动脉的影响。应用酶联免疫生化技术检测各组兔基底动脉血管平滑肌细胞膜蛋白激酶C（PKC）活性。结果经脑血管造影证实该剂量甲基强的松龙明显减轻实验性脑血管痉挛的严重程度，与对照组相比，PKC活性在大剂量甲基强的松龙治疗组没有明显提高。结论大剂量甲基强的松龙能够明显减轻脑血管痉挛程度，通过抑制血管平滑肌细胞来防治脑血管痉挛的发生发展。     

蛛网膜下腔出血大鼠基底动脉血小板源性生长因子的表达变化

目的探讨血小板源性生长因子(PDGF)在大鼠蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)血管壁的表达及关系。方法将30只大鼠按照枕大池二次注血的方法建立模型,然后分别于建立模型后的1 d、3 d、5 d、7 d、14 d将大鼠处死,取出基底动脉制作石蜡切片在光镜下观察。采用免疫组化法检测大鼠基底动脉血管壁PDGF的表达水平。结果模型组中PDGF在基底动脉血管壁上的表达,3 d和5 d组最明显,与脑血管痉挛程度的变化是一致的。结论通过枕大池二次注血能够成功的模拟SAH后CVS的发生。PDGF参与了SAH后CVS的过程,并可能起了重要的作用。     

ICAM-1 and VCAM-1 expression following aneurysmal subarachnoid hemorrhage and their possible role in the pathophysiology of subsequent ischemic deficits

BACKGROUND: The pathophysiology of ischemic cerebral lesions following aneurysmal subarachnoid hemorrhage (SAH) is poorly understood. There is growing evidence that inflammatory reactions could be involved in the pathogenesis of such delayed occurring ischemic lesions. The aim of this study was to evaluate adhesion molecules with regard to these lesions following SAH. METHODS: Serum and cerebrospinal fluid (CSF) samples were taken daily from 15 patients up to day 9 after SAH and evaluated for intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1). RESULTS: CSF and serum samples correlated well during nearly the whole time course (p < 0.0001). A secondary increase in ICAM-1 and VCAM-1 in the serum and CSF correlated with an increase in flow velocity in the transcranial Doppler (p > 0.0001 and p < 0.007) but not to a delayed lesion in the CT scan. CONCLUSION: We believe that inflammatory processes are involved in the pathogenesis of cerebral vasospasm but they might only be a part of a multifactorial pathogenesis.     

褪黑素对大鼠脑缺血时白细胞浸润和 ICAM-1表达的影响

目的研究褪黑素(melatonin,MT)对大鼠脑缺血损伤缺血区髓过氧化物酶(myeloperoxidase,MPO)的活性及细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)表达的影响。方法线栓法造成大鼠大脑中动脉栓塞制成脑缺血模型,手术分为正常对照组、缺血组、MT治疗组,MT治疗组缺血前30min给予MT (20mg/kg)腹腔注射。术后在相应时间点取缺血侧脑组织,用分光光度计检测MPO的活性,用免疫组化检测I- CAM-1蛋白的表达。结果与正常对照组比较,缺血组各时间点MPO的活性及ICAM-1蛋白的表达均升高,而与缺血组比较MT治疗组各时间点MPO的活性及ICAM-1蛋白的表达均降低。结论MT可能能通过降低MPO的活性即抑制白细胞的浸润和降低ICAM-1蛋白的表达,对缺血的脑组织产生保护作用。     

Soluble and cell surface ICAM-1 as markers for disease activity in multiple sclerosis

Objective - The intercellular adhesion molecule-1 (ICAM-1) is a member of the Ig supergene family. ICAM-1 is expressed on various cells like peripheral blood lymphocytes, endothelial cells or thymic cells and the cell surface form is supposed to be shed into a soluble form. The expression of ICAM-1 is induced by cytokines like Interleukin-1, TNF alpha or interferon gamma. The aim of the study was to investigate whether changes of cell surface and soluble ICAM-1 in the cerebrospinal fluid (CSF) and blood are indicative for disease activity in patients with multiple sclerosis (MS). Material and methods - In all patients with relapsing-remitting MS (relapse: n =31, remission: n = 11) and controls ( n = 13) the expression of cell surface ICAM-1 (c-ICAM-1) was determined by two colour flow cytometry. Soluble ICAM-1 (s-ICAM-1) was measured by ELISA. Follow-up examinations were done 3 months later. Results - In 31 patients with a current relapse we found significantly decreased expression levels of c-ICAM-1 on leukocytes in CSF ( P <0.001) and blood ( P <0.10), when compared to those 11 individuals experiencing remission. In contrast we observed significantly ( P <0.05) increased levels of s-ICAM-1 in CSF of patients with relapses. Comparing patients who had been in remission for more than 4 weeks ( n = ll) with remission lasting longer than 3 months ( n =28) we detected stable c-ICAM-1 expression on CD3 ⁺ T cells in blood. Conclusion - Our results demonstrate for the first time that c-ICAM-1 on CD3 ⁺ T-cells in CSF and blood is an activity marker in MS.     

细胞间黏附分子-1启动子甲基化水平与脑梗死的相关性

目的 评估细胞间黏附分子-1(Intercellular adhesion molecule-1,ICAM-1)基因启动子区甲基化水平与脑梗死的相关性。方法 对2017年9月-2018年9月于解放军第960医院就诊患者进行回顾性分析,根据诊断标准挑选152例脑梗死患者为病例组,同期于该院体检健康者 152 例为对照组,采用酶联免疫吸附法(Enzyme linked immunosorbent assay,ELISA)检测2组受试者外周血ICAM-1水平,使用荧光定量甲基化特异性聚合酶链反应(Quantitative Methylation Specific PCR,qMSP)法测定2组受试者ICAM-1甲基化程度。结果 病例组外周血ICAM-1水平显著高于对照组(T=20.27,P<0.001); 病例组ICAM-1基因甲基化程度显著低于对照组(Z=-3.158,P=0.002); 病例组亚组分析发现有吸烟史者甲基化程度更低(Z=-3.305,P=0.001); 2组ICAM-1甲基化程度均与ICAM-1水平呈显著负相关(病例组r=-0.756,P<0.001; 对照组r=-0.823,P<0.001)。结论 ICAM-1低甲基化通过促进ICAM-1的高表达,参与脑梗死的病理过程,可能通过调节基因表达水平来影响脑梗死的患病风险。ICAM-1启动子区甲基化程度升高可作为脑梗死的保护因素。     

Nonenzymatic derived lipid peroxide, 8-iso-PGF2 alpha, participates in the pathogenesis of delayed cerebral vasospasm in a canine SAH model

We studied whether 8-iso-PGF2alpha, nonenzymatic arachidonyl peroxide, participated in the pathogenesis of delayed vasospasm using a canine subarachnoid hemorrhage (SAH) model. Fourteen adult mongrel dogs were divided into two groups, two-hemorrhage SAH group (n = 8) and control group (n = 6). The contents of 8-iso-PGF2alpha in CSF, the basilar artery segment, and subarachnoid clot were measured by enzyme immunoassay kit. The CSF 8-iso-PGF2alpha content on Day 7 in the SAH group was 67.9+/-29.9 pg ml(-1) (n = 8), which was significantly higher than 27.1+/-13.8 (n = 8) on Day 0 in the SAH group, and 33.2+/-14.4 pg ml(-1) (n = 5) on Day 7 in the control group. The 8-iso-PGF2alpha content in the basilar artery segment with spasm on Day 7 in the SAH group was 13.5+/-1.9 pg mg(-1) wet weight (n = 8), significantly higher than 8.7+/-1.9 (n = 6) in the control group. The 8-iso-PGF2alpha content in subarachnoid clot was 1.7+/-1.4 ng g(-1) wet weight (n = 8). Significant elevation of the 8-iso-PGF2alpha contents in the CSF and the basilar artery segment occurred on Day 7 in the SAH group. The subarachnoid clot enclosed the basilar artery on Day 7, contained a considerable amount of 8-iso-PGF2alpha. These results suggested that 8-iso-PGF2alpha could play a crucial role in the pathogenesis of the delayed cerebral vasospasm.     

亚低温对局灶性脑缺血大鼠白细胞介素-6和细胞间黏附分子-1表达的影响

目的 探讨亚低温对局灶性脑缺血大鼠缺血局部细胞间黏附分子-1（ICAM-1）表达和血清白细胞介素-6（IL-6）含量的影响。方法 将30只体重在250～300&#8197;g的雌性SD大鼠随机分为实验组（n=15）和对照组（n=15），采用线栓法阻断大鼠一侧大脑中动脉制作局灶性脑缺血模型.制模成功后，实验组和对照组大鼠分别给予亚低温、常温处理，使其肛温保持在（33±1）℃和（37±0.5）℃。12&#8197;h后，自左室取血，断头取脑，检测缺血区ICAM-1阳性血管数目（免疫组化方法）和血清IL-6含量（免疫放射测定法）。结果 实验组缺血局部ICAM-1的表达明显低于对照组[（2.45±1.56）vs（18.27±2.45）个/HP，P<0.01］；对照组血清IL-6含量、神经功能缺陷评分明显高于实验组[（184±38）vs（124±41）ng/L），P<0.01；（2.13±0.35）vs（1.48±0.25），P<0.01]。结论 亚低温对脑缺血性损害的神经保护作用可能与降低IL-6含量和减少ICAM-1的表达有关。     

Time course of ICAM-1 expression and leukocyte subset infiltration in rat forebrain ischemia

The time course of ICAM-1 expression and leukocyte subset infiltration was studied in a model of CNS reperfusion injury in adult rats. Leukocyte adhesion and infiltration, mediated in part by intercellular adhesion molecule-1 (ICAM-1), appears to potentiate CNS reperfusion injury. The timing and relationship between ICAM-1 staining and leukocyte infiltration postglobal CNS ischemia is unknown. Reversible forebrain ischemia was produced in 32 adult Sprague-Dawley rats using the two-vessel occlusion model with histologic analysis performed at specific intervals postischemia: 1, 3, 6, 12 and 24h, 4 and 7 d, or sham-operated controls (n=4 each group). Monoclonal antibodies against ICAM-1 (1A29 and TM8), a specific granulocyte (PMN) (HIS48), and a specific monocyte/macrophage (M?)(ED1) were used. No specific leukocyte and only rare ICAM-1 vessel immunoreactivity was observed in sham controls. ICAM-1: Significant expression in microvessels beginning at 1 h with additional diffuse CA1 pyramidal layer staining beginning at 4 d. Leukocytes: NO PMN cells and rare M? identified at 6 and 12 h. By 24 h: moderate infiltrate in areas of ICAM-1 expression of PMN and M?. At 4 and 7 d: only M? accumulation, cellular morphology now similar to microglia. The results of this study indicate that early and persistent ICAM-1 expression occurs following CNS ischemia with associated leukocyte infiltration.     

Effect of melatonin on cerebral vasospasm following experimental subarachnoid hemorrhage

OBJECT: The current study was undertaken to determine whether melatonin therapy reverses vasospasm and prevents apoptosis by inhibiting lipid peroxidation in an experimental subarachnoid hemorrhage (SAH) model. MATERIALS AND METHODS: The rabbits were divided into four groups as follows: Group 1, SAH + melatonin (5 mg/kg/i.p. BID) simultaneously with SAH (n = 6); Group 2, SAH + melatonin (5 mg/kg/i.p. BID) treated 2 hours after SAH (n = 6); Group 3, control group (n = 4); Group 4, SAH only (n = 6). Light microscopic examinations of the basilar arteries were performed to demonstrate the pathophysiological changes of the arterial wall with hematoxylin- eosin. Apoptosis: Immunohistology using the ApopTag Peroxidase In Situ Apoptosis Detection Kit was used to demonstrate apoptosis in a cross section of basilary arteries. Apoptotic index was calculated as the number of the immunoreactive nuclei per total number of endothelial cells, and expressed as a percentage. RESULTS: The results of measurements of diameters of the vessels between groups were significantly different (p = 0.028). While basilar arteries of the SAH only group showed 57% constriction, Groups 1 and 2 were calculated as 33 and 26% constriction, respectively, compared with the control group (p < 0.05). And also Groups 1 and 2 showed significant protection of apoptosis compared with Group 4. The difference between the four groups was tested by Kruskal-Wallis test and the significance between the two groups was tested by Mann- Whitney U-test. CONCLUSION: Melatonin with its strong antioxidant effect can prevent SAH-induced vasospasm and apoptosis of endothelial cells of vessels.     

易卒中型肾血管性高血压大鼠血浆CAM-1和P-选择素的动态观察

目的动态观察易卒中型肾血管性高血压大鼠（RHRSP）血浆ICAM-1和P-选择素的活性改变。方法双肾双夹法制作RHRSP模型,分别于术前及术后2、4、6、8、10、12、16周取血,酶联免疫吸附测定（ELISA）法检测ICAM-1和P-选择素含量。结果4周始RHRSP血中ICAM-1和P-选择素含量逐渐增高,明显高于正常对照组,且ICAM-1和P-选择素呈正相关,ICAM-1和P-选择素均与大鼠收缩压密切相关。结论血浆中ICAM-1和P-选择素含量升高提示梗死或出血的危险性增加,可作为脑卒中的预测指标之一。