肢体远隔缺血预适应对脑梗死患者的临床预后及炎性因子表达水平的影响

目的 探讨肢体远隔缺血预适应(RIPC)对脑梗死患者的临床预后及血清炎性因子表达水平的影响。方法 收集脑梗死患者271例为研究对象,随机分为RIPC组和对照组,RIPC组患者给予肢体缺血预适应训练及药物治疗,对照组给予单纯药物治疗; 观察2组患者治疗28 d及6个月后的神经功能恢复情况,记录发病6个月内的脑缺血症状再发生情况,对比2组患者治疗28 d后血清中炎性因子的表达水平及患者治疗过程中发生的不良反应。结果 治疗28 d及6个月后RIPC组NIHSS评分改善优于对照组(P<0.05); RIPC组的脑缺血再发生率低于对照组(P<0.05); 治疗28 d后RIPC组血清炎性因子表达水平显著低于对照组(P<0.05); 2组患者的不良反应发生率比较无明显差异(P>0.05)。结论 肢体远隔缺血预适应可显著改善脑梗死患者的短期和长期预后,降低脑缺血再发率,减轻体内的炎症反应,是一种安全有效的治疗方法。     

远隔肢体缺血后适应对不同TOAST和OCSP分型脑梗死的影响

目的探讨远隔肢体缺血后适应(RIPostC)在脑梗死TOAST分型和OCSP分型中对不同亚型脑梗死短期预后的影响。方法按标准选取住院的急性脑梗死患者545例,在常规治疗的基础上进行远隔肢体缺血后适应干预,分别于住院时和治疗7d后进行NIHSS评分,计算好转率。同时对所有入组患者按标准进行TOAST分型和OCSP分型,比较每个分型中不同亚型的NIHSS评分好转率。结果 TOAST分型中大动脉粥样硬化型、心源性栓塞型、小动脉闭塞型、其他原因的卒中型和不明原因型亚型之间的NIHSS评分好转率差异无统计学意义(P0.05)。OCSP分型中,完全性前循环梗死和部分性前循环梗死亚型的NIHSS评分好转率优于腔隙性梗死和后循环梗死亚型,差异有统计学意义(P0.05)。结论远隔肢体缺血后适应在脑梗死TOAST分型和OCSP分型中,对不同亚型脑梗死短期预后产生不同的影响,提示RIPostC应在适宜的、有选择的脑梗死亚型中应用。     

远隔缺血后处理对脑缺血再灌注损伤内质网应激相关因子的影响

目的探讨远隔缺血后处理(RIPC)对大鼠局灶性脑缺血再灌注损伤的保护作用,并观察RIPC对缺血再灌注损伤后内质网应激相关蛋白CRT、GRP78和caspase-12表达的影响。方法用线栓法制作大鼠大脑中动脉闭塞(MCAO)局灶性脑缺血再灌注模型,并进行远隔缺血后处理。再灌注6 h、12 h、24 h后分别采用Western blot检测CRT、GRP78和Bcl-2蛋白表达情况;RT-PCR检测caspase-12和caspase-3 mRNA表达情况。结果缺血再灌注组(6 h、12 h和24 h)与假手术组相比,CRT、GRP78、caspase-12、Bcl-2和caspase-3表达均升高,差异具有显著性(P<0.05)。远隔缺血后处理组与缺血再灌注组相比,CRT、GRP78和Bcl-2表达升高(12 h和24 h),差异具有显著性(P<0.05);caspase-12和caspase-3表达降低(6 h、12 h和24 h),差异具有显著性(P<0.05)。结论远隔缺血后处理可以减轻大鼠局灶性脑缺血再灌注产生的损伤,其保护作用机制可能与内质网应激反应有关。     

益气活血利水颗粒对急性脑出血后脑水肿的临床疗效及对MMP-9的影响

目的评价益气活血利水颗粒治疗急性出血性卒中的临床疗效和安全性;观察益气活血利水颗粒对血清基质金属蛋白酶-9(matric metalloproteinase-9,MMP-9)的影响,揭示其作用机制,为益气活血利水颗粒的临床应用提供客观依据。方法采用随机、对照的临床研究方法将60例出血性卒中患者分为益气活血利水颗粒治疗组(治疗组)20例和常规治疗对照组(对照组)20例。对照组给予脑出血后神经内科常规处理。治疗组在常规治疗基础上,并于当天口服益气活血利水颗粒(由三九药业生产的中药颗粒剂,主要成分:黄芪颗粒20g,党参颗粒15g,泽泻颗粒20g,水蛭粉颗粒3g),每日分2次冲服,2组均治疗30d,随访半年。对治疗组脑出血后患者血清MMP-9的含量、脑水肿面积及血肿面积,神经功能缺损评分,生活活动能力评分情况进行统计。结果治疗后2组神经功能缺损评分、脑水肿面积,血肿面积及生活活动能力评分比较,治疗组优于对照组,差异有统计学意义(P0.05)。治疗后第3天和第7天治疗组血清MMP-9水平较对照组显著降低(P0.05)。2组均未出现明显不良反应,患者能够耐受,2组患者均完成试验,没有病例因为严重不良事件而中断治疗。结论益气活血利水颗粒能抑制急性出血性卒中微创术后脑水肿形成;促进血肿的吸收;促进急性出血性卒中微创术后患者的神经功能恢复。临床研究未发现明显不良反应及毒副作用,安全性好。     

缺血后处理通过水通道蛋白-4对大鼠缺血再灌注损伤的脑保护作用及其机制

目的研究缺血后处理(IP)对大鼠脑缺血再灌注损伤的保护作用,探讨各组大鼠水通道蛋白-4(AQP4)的表达。方法实验分组:48只雄性SD大鼠,随机分为3组(n=16)。假手术组;对照组:行单纯缺血再灌注;缺血后处理组:IP组。采用线栓法阻断大脑中动脉制备大鼠局灶性脑缺血再灌注模型。大鼠脑缺血再灌注后24 h进行神经行为学评分和脑梗死体积测定,干湿重法测定脑水含量。并行免疫组织化学方法检测海马CA_1区细胞凋亡及AQP4表达的变化并行统计学分析。结果与对照组比较,IP组神经行为学评分明显降低,脑梗死体积、脑组织中的水分含量明显减少(P 0. 05)。再灌注24 h后,对照组海马CA_1区AQP4、凋亡细胞24 h表达量明显增加,IP组与对照组之间比较有差异(P 0. 05)。结论 IP组脑神经行为学评分、脑含水量、脑梗死体积均明显降低,提示缺血后处理可通过抑制AQP4的表达减轻缺血再灌注损伤后的脑水肿,起到脑保护作用。     

盐酸纳美芬对大鼠颅脑损伤后脑水肿的影响

目的探讨盐酸纳美芬对大鼠颅脑损伤后脑水肿的影响。方法将54只Wistar大鼠按随机数字表法随机分为假手术组、颅脑损伤组及纳美芬治疗组。采用自由落体法建立大鼠颅脑损伤模型,分别于伤后6 h、1 d、3 d、7 d,采用干湿重法检测脑组织含水量,应用免疫组织化学法检测损伤脑组织中水通道蛋白4（AQP4）的表达。结果与假手术组比较,颅脑损伤组和纳美芬治疗组的脑组织含水量及AQP4水平明显升高（P〈0.05）;与颅脑损伤组相比,纳美芬治疗组脑组织含水量及AQP4表达水平明显降低（P〈0.05）。通过相关性分析发现,大鼠颅脑损伤后脑组织AQP4表达水平与脑含水量呈明显正相关性（r=0.676,P〈0.01）。结论盐酸纳美芬可能通过抑制AQP4表达,减轻脑水肿,发挥神经保护作用。     

水通道蛋白4：缺血后脑水肿的靶分子

卒中后1周内，脑水肿引发的水快速流动导致脑组织膨胀是引起患者死亡的主要原因之一。对这个过程治疗的困难主要还是因为其机制仍然不是很清楚。1988年第一个水通道蛋白的发现，似乎为认识和治疗缺血后脑水肿开辟了一条崭新的道路。水通道蛋白（aquaporin,AQP），又称水孔蛋白，是一个同源水通道蛋白家族，在许多上皮和内皮细胞丰富表达，参与液体的转运。目前已发现的AQP有11种，其分布有相对的组织特异性，主要存在于脑内并且研究较多的是AQP1和AQP4。     

舒血宁注射液对脑缺血再灌注大鼠缺血半暗带水通道蛋白1和水通道蛋白9表达的影响

目的观察舒血宁注射液对脑缺血再灌注大鼠额顶叶皮质缺血半暗带(IP)水通道蛋白1(AQP1)和水通道蛋白9(AQP9)表达的影响。方法线栓法制作局灶性脑缺血再灌注大鼠模型(MCAO),随机分为假手术组、模型组、舒血宁注射液防治组和川芎嗪药物对照组。脑缺血2h,分别再灌注1d、3d、7d,采用免疫组化SABC法检测AQP1和AQP9蛋白表达的阳性总面积和平均吸收光密度。结果在缺血侧额顶叶皮质IP,再灌注各时间点,模型组AQP1和AQP9蛋白表达的阳性总面积和平均吸收光密度显著高于假手术组(P0.05或P0.01),舒血宁注射液和川芎嗪药物对照组再灌注1d、3d、7d,AQP1和AQP9蛋白表达的阳性总面积和(或)平均吸收光密度显著低于模型组(P0.05或P0.01)。结论舒血宁注射液能通过抑制AQP1、AQP9蛋白表达,减轻脑水肿,抗脑缺血再灌注损伤。     

肢体缺血预处理对大鼠缺血再灌注海马HSP70表达的影响

热休克蛋白(heat shock proteins70,HSP70),是一种应激蛋白。研究显示,大鼠大脑中动脉阻断后,HSP70蛋白仅出现在梗死灶周围神经元,即在缺血半影区大量表达。在短暂缺血后死亡的海马CA1区锥体细胞中HSP70蓄积甚少,存活的齿状回颗粒细胞中有明显的HSP70积累。以上表明HSP70可能对脑缺血后神经细胞具有保护作用。我们以前研究结果证实肢体缺血预处理(limb ischemic preconditioning,LIP)可减轻脑缺血再灌注损伤。本实验运用免疫组化技术观察脑缺血再灌注前给予LIP海马HSP70的表达情况,探讨HSP70在LIP抗脑缺血再灌注损伤中的作用。…     

远隔缺血后适应通过调节反应性星形胶质细胞的可塑性改善小鼠缺血后脑组织的恢复

目的星形胶质细胞的可塑性改变可以在脑缺血急性期引起脑水肿,在缺血恢复期改变胶质瘢痕的形成。本实验研究远隔缺血后适应(RIPC)在脑缺血再灌注损伤中对星形胶质细胞可塑性调节。方法脑缺血被诱导通过C57小鼠大脑中动脉短暂性的阻断1 h,在再灌注即刻给予RIPC。结果 RIPC能降低小鼠脑缺血再灌注3 d、14 d大脑半球的水肿、梗死面积,减少脑萎缩,提高神经功能恢复和生存率。而且RIPC能调节星形胶质细胞亚型的比例,在小鼠脑缺血再灌注3 d和14 d,RIPC能降低缺血侧纤维型星形胶质细胞(GFAP)及增加原浆型星形胶质细胞(GS)的表达,并且能够下调GFAPα的水平和上调GFAPδ/GFAPα的比例来调节GFAP的亚型。结论RIPC治疗能调节反应性星形胶质细胞的可塑性,提高缺血后神经功能的恢复。     

The effect of arterial hypertension on focal ischemic edema

Summary The middle cerebral artery (MCA) of cats was occluded permanently for 24h to study the influence of arterial hypertension during the early phase of focal ischemia upon the development of edema and changes of the blood-brain barrier (BBB). In normotensive animals MCA occlusion results in a hemispheric weight increase of about 8% and marked water and electrolyte alterations in both the grey and white matter of the MCA territory. The RISA space increases mainly in the grey matter. Hypertension aggravates these changes significantly, whereby water and electrolyte changes in the grey matter are predominantly concerned, while there is a preferential increase of the RISA space in the white matter. It is suggested that arterial hypertension aggravates the ischemic edema and enhances a vasogenic type of edema in the white matter.

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Zusammenfassung Die Veränderungen des fokalen Hirnödems sowie der Bluthirnschrankenfunktion durch eine arterielle Hypertension wurde 24 Stunden nach permanentem Verschluß der Arteria cerebri media der Katze untersucht. Bei den normotensiven Kontrolltieren fanden sich eine Zunahme des Gewichtes der geschädigten Hemisphäre auf über 8% sowie signifikante Wasser- und Elektrolytveränderungen in grauer und weißer Substanz des Mediaversorgungsgebietes. Eine Zunahme des sog. RISA-Raumes betrifft bevorzugt die graue Substanz. Nach Hypertension nehmen die regionalen Wasser- und Elektrolytveränderungen signifikant zu, wobei besonders in der grauen Substanz die Wasser- und Elektrolytveränderungen auftreten, während in der weißen Substanz die Zunahme des RISA-Raumes betont ist. Aus den vorliegenden Befunden wird geschlossen, daß eine arterielle Hypertension das ischämisch-fokale Hirnödem verstärkt und besonders in der weißen Substanz das Bild eines vasogenen Ödemtyps aufweist.

     

Protective effect of delayed remote limb ischemic postconditioning: role of mitochondrial K(ATP) channels in a rat model of focal cerebral ischemic reperfusion injury

Delayed remote ischemic postconditioning (DRIPost) has been shown to protect the rat brain from ischemic injury. However, extremely short therapeutic time windows hinder its translational use and the mechanism of action remains elusive. Because opening of the mitochondria K(ATP) channel is crucial for cell apoptosis, we hypothesized that the neuroprotective effect of DRIPost may be associated with K(ATP) channels. In the present study, the neuroprotective effects of DRIPost were investigated using adult male Sprague-Dawley rats. Rats were exposed to 90 minutes of middle cerebral artery occlusion followed by 72 hours of reperfusion. Delayed remote ischemic postconditioning was performed with three cycles of bilateral femoral artery occlusion/reperfusion for 5 minutes at 3 or 6 hours after reperfusion. Neurologic deficit scores and infarct volumes were assessed, and cellular apoptosis was monitored by terminal deoxynucleotidyl transferase nick-end labeling. Our results showed that DRIPost applied at 6 hours after reperfusion exerted neuroprotective effects. The K(ATP) opener, diazoxide, protected rat brains from ischemic injury, while the K(ATP) blocker, 5-hydroxydecanote, reversed the neuroprotective effects of DRIPost. These findings indicate that DRIPost reduces focal cerebral ischemic injury and that the neuroprotective effects of DRIPost may be achieved through opening of K(ATP) channels.     

局灶脑缺血后胰岛素对c-fos、c-jun基因表达的影响

目的　观察胰岛素对局灶缺血性脑组织 c- fos、c- jun基因表达的影响。方法　 30只肾血管性高血压大鼠随机分 4组 ,在大脑中动脉闭塞 (MCAO)后即予胰岛素 (2 .1IU / kg,A组 )、葡萄糖和胰岛素 (2 .1IU / kg,B组 ;4.5 IU / kg,C组 )、生理盐水 (D组 ) ,采用原位杂交技术检测各组 MCAO后 3h脑组织 c- fos、c- jun基因的表达。结果　与其它组比较 ,A组血糖下降明显 (P<0 .0 1) ,MCAO后 1h、2 h、3h分别为 3.80± 0 .3mm ol/ L、3.6 8± 0 .3mm ol/L、3.6 9± 0 .3mm ol/ L。 c- fos m RNA被诱导出现于缺血侧皮层和尾壳核 ,C组 c- fos m RNA明显增加 (P<0 .0 1) ,皮层和尾壳核的灰度为 12 3.5 8± 8.72、141.0 8± 8.49;c- jun m RNA仅出现于缺血侧皮层 ,A、B、C组的 c- jun m RNA均增加明显 ,灰度分别为 171.82± 7.33、172 .5 6± 7.91、15 5 .0 9± 7.91,其中 C组的增加更为显著 (P<0 .0 1)。结论　胰岛素促进缺血脑组织 c- fos、c- jun基因表达可能是其神经保护作用机制之一。     

阿加曲班对大鼠脑出血模型脑水肿作用的研究

目的 探讨阿加曲班对大鼠脑出血模型脑水肿的作用.方法取雄性SD大鼠40只,体重300～350g,随机分为A、B、C、D、E组,每组8只.大鼠用10%水合氯醛(350mg/kg)腹腔注射麻醉后,经立体定向分别向右侧尾状核区注入生理盐水60μl、无肝素自体血50μl 生理盐水10μl、无肝素自体血50μl Argatroban生理盐水溶液(0.5μl/ml)10μl、含10U凝血酶生理盐水溶液50μl 生理盐水10μl、含IOU凝血酶生理盐水溶液50μl Ar-gatroban生理盐水溶液(0.5μg/ml)10μl.48h后,断头处死,测定脑含水量,HE染色,观察脑组织病理学变化.结果 C、E两组脑组织含水量降低,与B、D两组比较有明显差别(P<0.05);B组与D组比较脑组织含水量无明显差别(P>0.05).HE染色观察:B组可见血肿周围脑组织水肿,炎症细胞渗出,脑组织疏松,神经细胞呈气球样变、坏死;D组可见注射部位脑组织水肿,脑组织疏松,炎症细胞渗出;C、E两组上述改变减轻;A组未见血肿及局部明显病理学改变.结论阿加曲班可减轻凝血酶造成的脑水肿;可减轻大鼠脑出血后血肿周围脑组织脑水肿     

阿加曲班对大鼠脑出血后脑水肿作用的研究

目的探讨阿加曲班对大鼠脑出血模型脑水肿的作用。方法取雄性SD大鼠32只,体重300~350g,随机分为A、B、C、D,4组,每组8只。大鼠经立体定向分别向右侧尾状核区注入生理盐水60ul、无肝素自体血50ul+生理盐水10ul、生理盐水50ul+阿加曲班生理盐水溶液10ul、无肝素自体血50ul+阿加曲班生理盐水溶液10ul。实验大鼠于术后48小时断头处死,测定脑含水量,HE染色,观察脑组织学变化。结果D组脑组织含水量降低,与B组比较有明显差别(P<0.05);C组脑组织含水量与A组比较无明显差别(P>0.05);HE染色观察:B组可见血肿周围脑组织水肿,炎症细胞渗出,脑组织疏松;D组上述改变减轻;A、C两组未见明显病理学改变。结论阿加曲班可减轻大鼠脑出血后血肿周围脑组织脑水肿。     

Aging has a complex effect on a rat model of ischemic stroke

Stroke in humans is usually associated with advanced age. Nevertheless, almost all animal models of ischemic stroke are based on young animals. The present study was designed to assess the effect of age on the development of ischemic injury in a model of focal brain ischemia in rats. Two age groups of Wistar rats were used: young adult (3 months) and old (24-26 months). Under halothane anesthesia, polyethylene microspheres (50 microm in diameter) were injected into the left common carotid artery following a temporary occlusion of the external carotid artery. Sham-operated rats underwent the same procedure but were injected with an identical volume (100 microl) of saline only. Rats of both experimental groups displayed neurological impairment after surgery. However, contrary to expectation, the young rats were more affected than the old rats. Young rats displayed an abrupt 30% decrement in neurological functions in the first week and then showed a partial functional recovery into a 12% decrement from the second week on. Old rats developed the neurological impairment gradually over a 2-week period (6.3% in the first week and 11% in the second week and thereafter). One month later, rats were tested in a water maze task. Again, performance was more impaired in the young ischemic rats than in the old rats. Histological evaluation revealed more extensive neurological damage in young ischemic as compared to old rats. Thus, although increased age has a critical effect on the evolution of the neurological impairment following focal brain ischemia and stroke, its effects in the rat model were more pronounced in the young animals.     

Effects of fluid management on edema volume and midline shift in a rat model of ischemic stroke

BACKGROUND AND PURPOSE: The purpose of this study was to investigate the effects of fluid management on brain water content (BW) and midline shift (MLS) after a focal cerebral ischemic insult. METHODS: A suture model was used to induce focal cerebral ischemia for 90 minutes (n=44). The rats were randomly assigned to 3 groups 2. 5 hours after reperfusion: dehydration (n=24), control (n=8), or hydration (n=12). BW was obtained with the wet-dry weight method 24 hours after middle cerebral artery (MCA) occlusion. In addition, MRI were obtained (n=31) 24 hours after the onset of ischemia so that the ratio of hemispheric volumes ipsilateral (IH) and contralateral (CH) to the infarct and the extent of MLS could be obtained. RESULTS: Across the range from moderate dehydration to intravascular volume expansion with isotonic saline, BW of the IH increased linearly as a function of change in body weight (r(2)=0.89), whereas few changes in relation to body weight were observed in CH, indicating a preferential effect of fluid management on the infarcted hemisphere. Furthermore, the hemispheric volume ratio (IH/CH) and MLS also increased in relation to changes in body weight. However, paradoxical increases in BW, IH/CH, and extent of MLS were observed in comparison with controls when severe dehydration was produced with high-dose mannitol. CONCLUSIONS: Changes in ischemic BW by fluid management correlated closely with changes in body weight except when high-dose mannitol was used. Mannitol, as a dehydrating agent, may be associated with bimodal effects, with a high dose aggravating ischemic BW.