国产降纤酶对大鼠缺血/再灌注脑损伤的保护作用

目的观察3种国产降纤酶对大鼠缺血/再灌注脑损伤的保护作用。方法采用线栓法大鼠局灶性脑缺血/再灌注模型,观察3种国产降纤酶对缺血/再灌注不同时程动物脑梗死体积、血流量、神经功能缺损评分及梗死灶内肉眼出血率的影响。结果持续缺血3h,降纤酶治疗的各组动物脑梗死体积明显小于生理盐水对照组(P<0.05);缺血3h再灌注3h和72h,降纤酶治疗的各组动物脑梗死体积与生理盐水组相比无明显变化,但缺血3h再灌注6h和24h,降纤酶治疗的各组动物脑梗死体积比生理盐水组明显减少(P<0.05)。缺血3h再灌注6h、24h和72h,降纤酶治疗组动物脑血流量比生理盐水组明显增加(P<0.05)。但治疗组动物行为学评分较生理盐水组无相应改善,梗死灶内有肉眼出血的动物较生理盐水组多,但无统计意义。结论国产降纤酶能明显减小缺血/再灌注脑损伤动物的梗死体积和增加脑血流量,改善损伤后的低灌注状态,对脑组织有一定保护作用。     

人尿激肽原酶对局灶性脑缺血再灌注大鼠VEGF表达影响的研究

目的 探讨人尿激肽原酶对局灶性脑缺血再灌注大鼠脑组织血管内皮生长因子(VEGF)表达的影响.方法 采用随机数字表法将56只雄性SD大鼠分为假手术组(8只)、生理盐水组(24只)、人尿激肽原酶组(24只),其中生理盐水组、人尿激肽原酶组依据再灌注后不同取材时间又分为6 h,12 h,24 h,72 h,7 d五个亚组.采用线拴法制备大鼠局灶性脑缺血再灌注模型,采用神经功能评分、TTC染色、脑梗死体积测定、光镜检测等方法对不同组大鼠予以评价.采用免疫组化技术观察缺血再灌注不同时间点大鼠脑组织梗死中心区及半影区VEGF表达变化情况.结果 人尿激肽原酶组大鼠神经功能评分低于生理盐水组大鼠(P<0.05);24 h脑梗死体积测定,人尿激肽原酶组平均值为(53 261.96±7 326.75)μm3,生理盐水组平均值为(92 715.84±13 755.44)μm3,差异有统计学意义(P<0.05);人尿激肽原酶组VEGF表达在不同时间点均明显强于生理盐水组(P<0.05).结论 人尿激肽原酶能减轻脑缺血再灌注模型大鼠的神经功能损伤程度,减少脑梗死体积,促进VEGF的表达,具有脑缺血后神经保护作用.     

三七三醇皂苷对脑缺血再灌注大鼠的保护作用

目的 通过对局灶性脑缺血大鼠不同再灌注时段的动态观察．探讨三七三醇皂苷(PTS)对大鼠局灶性脑缺血／再灌注动物模型的神经行为学和脑梗死体积的保护作用。方法 采用改良的线栓法制备大脑中动脉阻塞(MACO)2h、再灌注不同时间段(3h、6h、12h、24h、48h、72h、7d)的大鼠短暂局灶性脑缺血模型。动物随机分假手术组、生理盐水对照组、三七三醇皂苷(PTS)组。用Zea Longa5分制评分和TTC染色法评价神经行为学和脑梗死体积。结果 神经行为学评分除72h组有明显改善外．其余各组与生理盐水对照组比较无显著性差异。脑梗死体积除再灌注3h、6h外．其余各组与生理盐水组比较差异均有显著性意义。结论 三七三醇皂苷对大鼠局灶性脑缺血及再灌注损伤有一定的保护作用。     

钙拮抗剂对大鼠脑缺血后血脑屏障通透性的影响

目的 研究钙离子拮抗剂对大鼠脑缺血再灌注后血脑屏障(BBB)通透性和脑梗死灶体积的影响. 方法 插线法制作大鼠脑缺血再灌注模型.缺血2 h后再灌注.将150只大鼠按随机数字表法分尼莫地平组和对照组,每组分再灌注6h、12h、24 h、48h、72 h五个时间段,再灌注后尼莫地平组和对照组立即分别腹腔注射尼莫地平和生理盐水2 mg/kg.每12小时注射一次,用甲酰胺荧光法及透射电镜观察不同时段BBB通透性破坏的情况,TTC染色后计算梗死灶体积百分比.结果 大鼠脑缺血再灌注后BBB通透性和梗死灶体积百分比随时间延长逐渐增加.且BBB通透性的增加呈现两个高峰,第一个高峰在再灌注后12 h,第二个高峰在再灌注后48 h.尼莫地平组BBB通透性及脑梗死灶体积百分比的增加均较对照组明显,差异有统计学意义(P<0.05). 结论 脑缺血再灌注增加BBB的通透性和脑梗死灶体积百分比.再灌注后给予尼莫地平可加重这些病理变化.     

低分子肝素对大鼠脑缺血后活化的小胶质细胞和巨噬细胞及TNF-α表达的影响

目的研究低分子肝素(LMWH)对大鼠局灶性脑缺血再灌注后活化的小胶质细胞和巨噬细胞及TNF-α表达的影响。方法建立大鼠大脑中动脉缺血再灌注模型,并将大鼠随机分为假手术组、对照组、LMWH治疗组,对照组及LMWH治疗组分别于脑缺血2h再灌注3、24、48、72h处死;应用免疫荧光染色检测脑组织中活化的小胶质细胞和巨噬细胞数量变化,应用免疫组化染色观察TNF-α阳性细胞。结果治疗组与对照组比较24、48、72h组梗死灶周围活化的小胶质细胞和巨噬细胞及TNF-α阳性细胞明显减少(P<0.05)。结论LMWH对大鼠脑缺血再灌注后梗塞灶周围活化的小胶质细胞和巨噬细胞及TNF-α表达有明显抑制作用。     

大鼠脑缺血再灌注后梗死体积的动态变化

目的观察大鼠大脑中动脉缺血再灌注后梗死灶体积的变化规律。方法线栓法制作大鼠局灶性脑缺血再灌注模型,观察脑缺血2h再灌注3h、24h、3d、7d、14d及21d后的神经功能缺损评分及2％氯化三苯基四氮唑(TTC)标记的梗死体积。结果缺血2h再灌注3h组已经出现较明显的梗死灶(梗死体积占前脑体积14．4％),再灌注24h组梗死体积最大(24．3％),显著大于再灌注3h、7d、14d、21d组(P<0．05)。再灌注3d组梗死灶仍较大 (23．8％),再灌注7d组梗死体积缩小(5．0％),再灌注14d组梗死灶进一步缩小(1．2％),再灌注21d组梗死灶基本消失(0．2％)。大鼠神经功能缺损评分与梗死体积之间呈显著相关(r=0．61,P<0．01)。结论脑缺血再灌注后梗死体积于24h达最大,21d时基本消失。脑缺血再灌注后神经功能缺损评分与梗死体积之间显著相关。     

腺病毒介导kallikrein基因过度表达对脑梗死周边区细胞凋亡的作用

目的 探讨激肽释放酶(kallikrein)对脑梗死周边区域缺血神经细胞凋亡的作用.方法 建立大鼠大脑中动脉闭塞(MCAO)模型,将造模成功的30只大鼠采用随机数字表法分为以下几组:A组,空白对照组;B组,注射生理盐水;C组,注射pAdCMV-人类组织激肽释放酶(HTK):每组10只.观察每组大鼠处理前后神经功能损害评分(NSS),用免疫组化技术检测外源性HTK的表达,并通过TUNEL染色、RT-PCR检测细胞凋亡及凋亡因子bcl-2、bax、caspase-3 mRNA水平.结果 治疗后24h在C组中可见HTK表达阳性细胞并逐渐增多,72h后达高峰.与B组以及A组相比差异有统计学意义(112±6.1、68±4.2、59±3.9,P<0.05).治疗后72h,C组大鼠NSS神经功能评分明显低于B组以及A组(6.70±0.16、8.13±0.16、7.93±0.20,P<0.05);治疗后7 d差异更明显(5.14±0.18、7.82±0.14、7.91±0.10,P<0.01).凋亡神经细胞集中于梗死周边区,治疗后72h及7d时,C组平均TUNEL阳性细胞数较B组与A组明显减少(72 h:10.1±0.9、16.7±1.1、20.4±0.8;7 d:15.2±1.2、33.6±1.3、28.8±1.7,P<0.05).与B、A组比较,C组中bc1.2 mRNA水平增高不明显(P>0.05);治疗后72 h及7 d,bax与caspase.3 mRNA水平则明显降低(P<0.05). 结论 Kallikrein可能通过减少凋亡因子bax、caspase-3的表达,抑制脑梗死周边区域的神经细胞凋亡,从而达到保护缺血神经细胞,改善神经功能的作用.     

糖尿病大鼠及局灶缺血再灌注模型的探讨

目的探讨一种比较简易的糖尿病大鼠局灶性脑缺血/再灌注模型制备方法并比较分析影响模型制备成功的因素。方法70只体质量为180~220g的雄性SD大鼠禁食12h后,于腹腔内一次性注射链脲霉素60mg/kg,选择体质量为280~320g血糖水平16.7~25.6mmol/L的成模实验性慢性糖尿病大鼠制作脑缺血再灌注模型;另选择55只体质量为280~320g的同种同月龄雄性大鼠制作单纯脑缺血再灌注模型作为对照;采用ZeaLonga线栓改进法,从rCBF、神经功能缺陷评分以及梗死灶体积等三个方面进行对比研究。比较2组大鼠模型制备成功率并分析2组大鼠模型制备失败及死亡原因。结果脑缺血再灌注模型制备:单纯脑缺血再灌注组大鼠神经功能评分低于糖尿病脑缺血再灌注组(P<0.01),2组模型制备成功后,大鼠脑组织的血流量在6h后进行性降低,可能与再灌注后的48h内脑组织水肿进行性加重有关。糖尿病组脑梗死灶体积在24、48h时明显比正常组大鼠严重(P<0.05)。结论此项研究为探索糖尿病与脑血管病变的关联提供了一种较为理想的动物模型。     

阿托伐他汀对大鼠脑缺血再灌注后梗死灶周围中性粒细胞浸润和核转录因子-κB表达水平的影响

目的 观察阿托伐他汀对大鼠脑缺血再灌注后梗死灶周围中性粒细胞浸润以及核转录因子-κB表达水平的影响。方法 采用常规尼龙线栓法制备SD大鼠脑缺血再灌注模型,并将大鼠随机分为假手术组、大脑中动脉阻断再灌注(Middle cerebral artery occlusion/reperfusion,MCAO/R)(对照)组和MCAO/R阿托伐他汀(治疗)组; 对照组和治疗组分别于脑缺血2 h再灌注3 h处死; 采用TTC染色测定脑梗死体积; 应用HE染色检查梗死灶周围中性粒细胞浸润; 应用免疫组化染色观察核转录因子-κB(Nuclear factor-κB,NF-κB)活性水平。结果 与对照组比较,治疗组脑梗死体积减小(P<0.01); 与对照组比较,治疗组再灌注3 h后梗死灶周围中性粒细胞明显减少(P<0.01); 与对照组比较,治疗组的脑组织NF-κB活性降低(P<0.01)。结论 阿托伐他汀对大鼠脑缺血再灌注后梗死灶周围中性粒细胞浸润有明显抑制作用; 阿托伐他汀可抑制大鼠脑缺血再灌注过程中的炎症反应,其机制可能与抑制NF-κB的激活有关。     

低分子肝素对大鼠脑缺血后活化的小胶质细胞和巨噬细胞及TNF-α仅表达的影响

目的 研究低分子肝素（LMWH）对大鼠局灶性脑缺血再灌注后活化的小胶质细胞和巨噬细胞及TNF-α表达的影响。方法 建立大鼠大脑中动脉缺血再灌注模型，并将大鼠随机分为假手术组、对照组、LMWH治疗组，对照组及LMWH治疗组分别于脑缺血2 h再灌注3、24、48、72 h处死；应用免疫荧光染色检测脑组织中活化的小胶质细胞和巨噬细胞数量变化，应用免疫组化染色观察TNF-α阳性细胞。结果 治疗组与对照组比较24、48、72 h组梗死灶周围活化的小胶质细胞和巨噬细胞及TNF-α 阳性细胞明显减少（P＜0．05）。结论 LMWH对大鼠脑缺血再灌注后梗塞灶周围活化的小胶质细胞和巨噬细胞及TNF-α表达有明显抑制作用。     

Effects of kallikrein gene transfer on penumbral microvascular proliferation and on regional cerebral blood flow following cerebral ischemia/reperfusion injury

BACKGROUND： Recent findings have demonstrated that the kallikrein-kinin system （KKS） participates in the pathological process of cerebral ischemia/reperfusion injury. Kallikrein gene transfer exhibits neural protective effects following cerebral infarction. OBJECTIVE： To observe the effects of kallikrein gene transfer on vascular proliferation in the peripheral infarct focus and on regional cerebral blood flow （rCBF） following cerebral ischemia/reperfusion injury. DESIGN, TIME AND SETTING： The completely randomized, controlled experiment was performed at the Lin Baixin Laboratory Center, the Second Affiliated Hospital of Sun Yat-sun University between September 2007 and April 2008. MATERIALS： pUCI9-HTK plasmid was constructed and maintained in the Laboratory for Neurology, the Second Affiliated Hospital of Sun Yat-sen University, China. Mouse anti-human kallikrein 1 monoclonal antibody was purchased from R＆D Systems, USA. METHODS Ninety healthy, male, Sprague Dawley rats were used. Middle cerebral artery occlusion （MCAO） was established in all rats to induce cerebral ischemia/reperfusion injury. Following MCAO establishment, all rats were randomly divided into three groups （n = 30）： blank control, saline, and pAdCMV-HTK. The saline and pAdCMV-HTK groups were stereotactically micro-injected with 5μL of physiological saline or with pAdCMV-HTK [multiplicity of infection （MOI） = 20], respectively, into the ischemic penumbra. In the blank control group, only sham injection was performed. MAIN OUTCOME MEASURES： At 12, 24, and 72 hours after treatment, cerebral infarction volume was measured by 2, 3, 5-triphenyltetrazolium chloride （TTC） staining. Exogenous HTK expression, as well as regional vascular endothelial growth factor （VEGF） expression, was detected by immunohistochemistry. rCBF was examined by 14C-iodoantipyrine micro tracing. In addition, neurological severity score （NSS） was performed. Higher scores indicated more severe neurological deficits. RESULTS： NSS res     

厄贝沙坦对大鼠局灶性脑缺血再灌注后炎症反应的影响

目的观察厄贝沙坦对大鼠局灶性脑缺血再灌注后脑内及外周炎症反应的影响。方法采用改良Longa方法制备大鼠大脑中动脉阻塞(middle cerebralartery occlusion,MCAO)模型,于缺血90min再灌注后24h和72h进行梗死体积的测量,采用免疫组化和ELISA方法测量脑内和外周血的粘附分子。结果厄贝沙坦可以显著减少局灶性脑缺血再灌注后24h和72h的梗死体积(均P<0.01),改善神经功能(均P<0.01);降低脑内ICAM-1、VCAM-1的表达及其外周血浆中可溶性的形式sICAM-1、sVCAM-1蛋白的水平(均P<0.05)。结论厄贝沙坦可以降低粘附分子的表达,减少梗死体积,改善神经功能,对脑缺血再灌注起保护作用。     

姜黄素对大脑中动脉缺血模型大鼠的神经保护作用及机制研究

目的 通过观察姜黄素（curcumin）对Notch 1及NF-κB表达的影响及脑含水量和梗死体积的变化,探 讨其对大脑中动脉梗死（middle cerebral artery occlusion,MCAO）模型大鼠的神经保护作用及机制。 方法 采用成年健康雄性Sprague-Dawl ey大鼠93只,随机分为假手术组（sham）,溶剂对照组（vehi cl econtrol ）,姜黄素组（CUR）。MCAO术后立即腹腔注射姜黄素溶液（80 mg/kg）,溶剂对照组及假手术 组给予同体积含0.5 mol/L NaOH的0.01 PBS。根据不同时间点每组分为对照、3 h、6 h、12 h、24 h、48 h、 72 h共7个亚组,分别在相应时间点进行神经功能学评分,2～4分者纳入实验组。归组后将动物断头 处死,留取病变侧脑组织利用免疫组织化学法及Western blot观察Notch 1及NF-κB的表达。各组仅取 48 h一个时间点进行脑含水量测定及2%的2,3,5-三苯基四唑氮红（triphenyltetrazolium chloride,TTC） 染色观测梗死体积。 结果 CUR组降低Notch 1和NF-κB的表达,这种抑制效果至少持续至MCAO后72 h（P＜0.05）。与 vehicle-control组相比,CUR组在MCAO后48 h时即可显著改善神经功能缺损（P＜0.05）,减少脑含水量 （[ 80.42±9.00）% vs（83.71±7.00）%（P＜0.05）]及梗死体积（[ 40.08±3.66）% vs（28.94±6.20）% （P＜0.05）]。 结论 姜黄素干预后,MCAO模型病变脑组织含水量降低,梗死体积减小,Notch 1和NF-κB表达水平 同步下调,推测姜黄素有脑保护作用,姜黄素可能通过抑制Notch 1和NF-κB的表达对缺血性脑组织 起到脑保护作用。     

Reduction of ischemic brain injury by anti-P-selectin monoclonal antibody after permanent middle cerebral artery occlusion in rat.

The selectin family of adhesion molecules is involved in adhesion of leukocyte to microcirculatory system and the transmigration into brain parenchyma. Although the role of P-selectin may be important in the pathogenesis of brain ischemia, a possible protective effect on ischemic brain injury by blocking P-selectin has not been reported. We have examined the effects of a novel anti-P-selectin antibody on ischemic brain injury after 24 h of permanent middle cerebral artery occlusion (MCAO) in rat. Male Wistar rats were subjected to MCAO by an insertion of a silicone rubber cylinder for 24 h. Anti-rat P-selectin monoclonal antibody, ARP 2-4, was injected intravenously at a dose of 1 mg kg-1 at 5 min before the induction of MCAO. Control animals received the same volume of vehicle solution. Regional cerebral blood flow (rCBF) was measured immediately after and at 8 h of MCAO. At decapitation of rats at 24 h of permanent MCAO, infarct size was compared between the antibody and vehicle treated group. In addition, immunohistochemistry for leukocyte infiltration and HSP72, and histochemistry for TUNEL were also, compared. Pretreatment with ARP 2-4 improved rCBF at 8 h of MCAO (55.4% +/- 11.7% of control, n = 5) as compared to vehicle group (24.2% +/- 11.8%, n = 5, p < 0.02). Although leukocyte infiltration was not normally detected by monoclonal antibodies for CD11a and CD18, it became remarkably evident at 1 day of MCAO. Although HSP72 and TUNEL were not also detected in sham control brains, they were induced in neurons of the MCA area at 1 day of MCAO. Treatment with ARP 2-4 significantly reduced the numbers of leukocyte and neurons with positive HSP72 and TUNEL stainings. These results demonstrated that an administration of a monoclonal antibody against P-selectin improved rCBF, and attenuated infarct size that was associated with reduction of leukocyte infiltration. Furthermore, treatment with the antibody reduced both HSP72 and TUNEL stainings. These data suggest an important role of P-selectin in ischemic brain damage, and a future therapeutic potential to human stroke patients.     

依达拉奉对局灶性脑缺血大鼠神经功能恢复及海马病理形态学改变的影响

目的 探讨大鼠局灶性脑缺血后神经功能恢复及海马病理形态学改变,评价依达拉奉的干预作用.方法 126只雄性SD大鼠随机分为3组:假手术组(A组,6只),生理盐水组(B组,MCAO后6h、12h、24h、48h、72h、5d、7d7时点,各6只),依达拉奉处理组(C组,同B组),术后即刻予以依达拉奉干预.行神经功能缺损评分测定;TTC染色观察脑梗死体积改变;用HE染色方法观察病理形态学改变.结果 术后进行神经缺损评分发现,由于麻醉和手术创伤的影响,假手术组术后出现6～24h神经功能减退.与假手术组相比,在6～24h时间段盐水组神经功能下降明显(P＜0.05).依达拉奉组神经功能明显好于盐水组(P＜0.05).术后7d盐水组和依达拉奉组神经功能基本恢复.同时,在依达拉奉组和盐水组中,缺血24h脑梗死体积最大;与盐水组相比,术后6～24h依达拉奉组脑梗死体积明显减小(P＜0.05).HE染色显示术后6h在脑缺血区神经元细胞无明显改变,6h后缺血区脑组织逐渐出现肿胀与坏死;在依达拉奉组,脑水肿和神经元坏死病理损害明显较轻,在假手术组,脑组织无明显改变.结论 依达拉奉具有改善神经功能缺损、缩小脑梗死体积和减轻缺血性病理损害程度的作用;研究还提示依达拉奉对缺血性脑卒中早期神经功能恢复具有十分重要的临床意义.     

头孢曲松钠对大鼠脑缺血损伤的保护作用及其机制

目的 探讨在大鼠局灶性脑缺血模型中应用头孢曲松钠对脑缺血损伤的保护作用及其相关机制.方法 制备Wistar大鼠局灶性脑缺血模型,并按随机数字表法分为单纯缺血组(MCAO组)、头孢曲松钠治疗组(MCAO+CTX组)和盐水对照组,其中MCAO+CTX组为缺血90min时给予头孢曲松钠200 mg/kg.缺血后24 h、48 h、7 d时对各组大鼠进行神经行为学评分和脑水肿程度测定,同时比较各组大鼠皮层和海马谷氨酸转运体功能的差异.结果 随着缺血时间延长,各组大鼠神经行为学评分逐渐提高;脑水肿在缺血后24 h、48 h时逐渐加重,至7 d时已逐渐消退.与MCAO组比较,各时间点MCAO+CTX组大鼠神经行为学评分明显提高,脑水肿程度明显减轻,伤侧皮层及海马谷氨酸转运体功能明显增强,差异均有统计学意义(P＜0.05).结论 头孢曲松钠对大鼠局灶性脑缺血损伤具有保护作用,其机制可能与增强谷氨酸转运体功能从而减轻谷氨酸神经毒性作用有关.

Abstract：

Objective To explore the neuroprotective effect of ceftriaxone on cerebral ischemia injury in rats with focal cerebral ischemia and its possible mechanism. Methods Focal cerebral ischemic models were established in Wistar rats and randomly divided into ischemic group (performed middle cerebral artery occlusion [MCAO]), ceftriaxone (CTX) therapy group (given CTX at a dosage of 200 mg/kg 90 min after MCAO) and control group (given physiological saline only). Twenty-four and 48 h, and 7 d after MCAO, neurological behaviors and cerebral edema level were evaluated in these 3 groups;glutamate transporter function in the cortex and hippocampus of rats was compared between each 2 groups. Results With time extended, neurological behaviors scores were obviously elevated in every group;and cerebral edema became worse at 24 and 48 h and decreased 7 d after MCAO. As compared with that in the ischemic group, glutamate transporter function, level of edema and neurological behaviors scores in cortex and hippocampus of rats in the CTX therapy group were statistically increased at different ischemic time points (P<0.05). Conclusion Ceftriaxone has a neuroprotective effect against focal cerebral ischemia in rats, which may relate to increased glutamate transporter function and reduced glutamate neurotoxicity.     

脑心通对大鼠局灶性脑缺血脑组织补体C3的影响

目的:探讨脑心通对大鼠局灶性脑缺血损伤后脑组织补体C3表达的影响。方法:参照longa等方法制成大鼠局灶性大脑中动脉阻断(MCAO)模型,将大鼠分为假手术组、生理盐水组、大剂量脑心通组、中剂量脑心通组、小剂量脑心通组,分别给予不同剂量药物,于术后6h,24h,48h,72h和7d进行神经功能评分(NDS),处死后取脑组织观察脑含水量的变化,病理切片HE染色观察组织病理学改变,免疫组织化学的方法测定补体C3的动态变化。结果:脑心通治疗组的神经功能评分均降低,病理损伤均减轻,大、中剂量脑心通组治疗后脑水肿明显减轻(P<0.05),补体C3的表达明显减少(P<0.05)。结论:脑心通可能通过减轻脑水肿,降低脑内补体C3的表达对大鼠缺血脑组织损伤产生保护作用。     

Reduction of ischemic brain injury by anti-P-selectin monoclonal antibody after permanent middle cerebral artery occlusion in rat

Abstract

The selectin family of adhesion molecules is involved in adhesion of leukocyte to microcirculatory system and the transmigration into brain parenchyma. Although the role of P-selectin may be important in the pathogenesis of brain ischemia, a possible protective effect on ischemic brain injury by blocking P-selectin has not been reported. We have examined the effects of a novel anti-P-selectin antibody on ischemic brain injury after 24 h of permanent middle cerebral artery occlusion (MCAO) in rat. Male Wistar rats were subjected to MCAO by an insertion of a silicone rubber cylinder for 24 h. Anti-rat P-selectin monoclonal antibody, ARP 2-4 was injected intravenously at a dose of 1 mg kg^–1 at 5 min before the induction of MCAO. Control animals received the same volume of vehicle solution. Regional cerebral blood flow (rCBF) was measured immediately after and at 8 h of MCAO. At decapitation of rats at 24 h of permanent MCAO, infarct size was compared between the antibody and vehicle treated group. In addition, immunohisto- chemistry for leukocyte infiltration and HSP72, and histochemistry for TUNEL were also compared. Pretreatment with ARP 2-4 improved rCBF at 8 h of MCAO (55.4% ± 11.7% of control, n = 5) as compared to vehicle group (24.2% ± 77.8%, n = 5, p < 0.02). Although leukocyte infiltration was not normally detected by monoclonal antibodies for CD11a and CD18, it became remarkably evident at 7 day of MCAO. Although HSP72 and TUNEL were not also detected in sham control brains, they were induced in neurons of the MCA area at 7 day of MCAO. Treatment with ARP 2-4 significantly reduced the numbers of leukocyte and neurons with positive HSP72 and TUNEL stainings. These results demonstrated that an administration of a monoclonal antibody against P-selectin improved rCBF, and attenuated infarct size that was associated with reduction of leukocyte infiltration. Furthermore, treatment with the antibody reduced both HSP72 and TUNEL stainings. These data suggest an important role of P-selectin in ischemic brain damage, and a future therapeutic potential to human stroke patients. [Neurol Res 1999; 21: 269-276]