经鼻给予TGFβ1对癫痫持续状态大鼠海马神经元的保护作用

目的探讨经鼻腔给予TGFβ1(transforming growth factor beta1,TGFβ1)对氯化锂-匹罗卡品所致癫痫持续状态(status epilepticus,SE)大鼠海马神经元的保护作用及其潜在的机制。方法健康雄性SD大鼠60只,随机分为转化生长因子(TGF)组、匹罗卡品(Pilo)组和正常对照组(control)。建立氯化锂-匹罗卡品癫痫持续状态模型。应用TUNEL染色、Fluoro-Jade B(FJB)荧光染色法分别观察各组大鼠海马神经元的原位凋亡及变性死亡情况。采用免疫组化方法检测凋亡相关基因caspase-3的蛋白表达。结果 SE后24h、48h、72h,TGF组大鼠海马FJB、TUNEL、caspase-3阳性细胞均较Pilo组显著减少(P<0.05);72h最为明显(P<0.01)。结论经鼻(IN)给予TGFβ1可以显著抑制或减轻癫痫持续状态大鼠海马神经元的变性与凋亡,从而发挥神经保护作用。其潜在的神经保护机制可能涉及下调caspase-3蛋白表达。     

褪黑素对癫(癎)大鼠海马神经元凋亡及半胱氨酸蛋白酶-3表达的影响

目的观察褪黑素（Mel）对癫癇大鼠海马神经元凋亡及半胱氨酸蛋白酶（caspase）-3表达的影响。方法采用匹罗卡品（Pilo）制作大鼠癫癇持续状态（SE）模型,随机分为Pilo组、Mel组和对照组,用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL）染色和免疫组化技术检测大鼠海马神经元凋亡数和caspase-3的表达,并与对照组比较。结果SE后6h,Pilo组开始出现少量TUNEL阳性细胞;SE后72h,达到高峰;SE后7d,TUNEL阳性细胞开始减少。SE后6h,Pilo组大鼠海马caspase-3阳性细胞数增多,主要集中于CA1和CA3区;SE后48h,达到高峰;SE后72h,阳性细胞数开始减少;SE后7d,caspase-3表达基本恢复正常。Mel组各时间点大鼠海马TUNEL阳性细胞数和caspase-3表达均明显低于Pilo组大鼠（均P〈0.01）。结论Mel可减少癫癇大鼠海马神经元凋亡,抑制caspase-3的表达,起到神经保护作用。     

右美托咪定调节MAPK/ERK-CREB通路对大鼠海马神经元凋亡的保护作用

目的探讨右美托咪定(DEX)调节MAPK/ERK-CREB通路对大鼠海马神经元凋亡的保护作用。方法通过腹腔注射氯化锂-毛果芸香碱构建癫痫持续状态(SE)大鼠模型,并随机分为4组,每组各10只。SE+DEX组在SE模型构建成功后腹腔注射DEX 1μmol/L,阳性对照组腹腔注射1μmol/L苯巴比妥,药物干预24 h后,通过Nissl法和TUNEL法检测大鼠海马神经元损伤及凋亡情况,Western blot法检测大鼠海马组织中MAPK、p ERK、p CREB蛋白和凋亡相关蛋白caspase-3、Bcl-2、Bax表达。结果与正常对照组相比,SE、阳性对照组、SE+DEX组大鼠Racine分值显著增加(P 0. 05),大鼠海马神经元数、Bcl-2蛋白表达量显著减少(P 0. 05),棕褐色TUNEL阳性细胞数、MAPK、p-ERK、p-CREB、caspase-3、Bax、Bax/Bcl-2蛋白表达量显著增加(P 0. 05)。与SE组相比,阳性对照组、SE+DEX组大鼠Racine分值显著降低(P 0. 05),大鼠海马神经元数、Bcl-2蛋白表达量显著增加(P 0. 05),棕褐色TUNEL阳性细胞、MAPK、p-ERK、p-CREB、caspase-3、Bax、Bax/Bcl-2蛋白表达量显著减少(P 0. 05)。结论 DEX可能通过抑制MAPK/ERK-CREB通路抑制海马神经元凋亡对其有保护作用。     

槲皮素对大鼠癫痫持续状态后海马XIAP mRNA与蛋白表达的影响

目的研究大鼠癫痫持续状态(SE)后海马组织XIAP的表达变化及槲皮素对其表达的影响。方法建立氯化锂-匹罗卡品致痫大鼠SE模型,应用免疫组化和RT-PCR方法检测XIAP与caspase-3蛋白以及XIAP mRNA的表达。结果海马CA3区XIAP蛋白在SE后2 h(0.5503±0.0172)起逐渐增加,8 h(0.6221±0.0238)达高峰,与对照组比较(0.1507±0.0165),差异有统计学意义(P<0.01)。海马CA3区caspase-3活性蛋白在对照组未见明显表达,在SE后4~72 h明显增多。与对照组比较,SE组海马XIAP mRNA表达水平在2~8 h增加(P<0.01)。与SE组比较,槲皮素组海马XIAP mRNA表达水平及CA3区XIAP蛋白表达在8 h、24 h高于SE组(P<0.01),CA3区caspase-3活性蛋白表达在8 h、24 h、72 h低于SE组(P<0.01)。结论XIAP可能参与了SE后神经元凋亡的调控过程,槲皮素可以提高SE后海马神经元XIAP的表达。     

颞叶癫痫大鼠海马XIAP相关因子-1蛋白表达变化研究

目的动态观察颞叶癫痫大鼠海马神经元XIAP相关因子-1(XAF1)的表达变化,探讨其在癫痫发生发展中的作用。方法清洁级SD雄性成年大鼠36只,随机分为正常对照组、致痫(SE)后3h、6h、12h、24h、72h组。用氯化锂(LiCl)和匹罗卡品(PILO)制备癫痫动物模型,应用免疫组化染色技术检测致痫后各时间点XAF1蛋白表达情况。结果对照组海马CA1、CA3区神经元XAF1蛋白表达极少,SE组CA1、CA3区XAF1蛋白表达在3～24h表达逐渐增高,72h表达有所下降,但仍高于对照组,各时间点两组间差异均有统计学意义(P0.01)。结论SE大鼠海马神经元XAF1表达增高,它可能参与了SE后脑神经元凋亡的调控。     

新型GLP-1/GIP双受体激动剂DA3-CH对大鼠癫痫持续状态后海马神经元凋亡的影响

目的探索新型胰高血糖素样肽-1/葡萄糖依赖性促胰岛素样肽(GLP-1/GIP)双受体激动剂DA3-CH对匹罗卡品诱导的大鼠癫痫持续状态(SE)后海马神经元凋亡的影响,进而探讨其神经保护作用。方法将108只体质量为250～300g的健康成年雄性SD大鼠随机分为正常对照组(NS组,n=12)、癫痫持续状态模型组(SE组,n=48)、DA3-CH干预组(SE+DA3组,n=48),SE组和SE+DA3组分别据造模成功后观察指标的不同时间点(SE后12h、1d、3d、7d)分为4个亚组(每个亚组n=12)。腹腔注射匹罗卡品(30mg·kg~(-1))诱导SE模型,造模成功后,SE+DA3组大鼠每日进食前给予腹腔注射DA3-CH(101nmoL·kg~(-1)),NS组及SE组大鼠给予等体积生理盐水。各亚组分别于SE后12h、1d、3d、7d检测空腹尾静脉血糖水平后处死取脑,采用免疫组化和Western blot方法检测海马中凋亡相关蛋白Bax、Bcl-2及神经元核抗原NeuN的表达水平。结果①NS组、SE组、SE+DA3组各亚组大鼠之间的空腹尾静脉血糖水平差异无统计学意义(P>0.05);②免疫组化与Western blot结果显示,NS组、SE组、SE+DA3组各亚组大鼠之间海马Bax、Bcl-2、NeuN表达水平差异均有统计学意义(P<0.001);与NS组相比,SE组大鼠海马Bax、Bcl-2表达水平升高,NeuN表达水平降低;与SE组相比,SE+DA3组大鼠海马Bax表达水平降低,Bcl-2、NeuN表达水平升高。结论新型GLP-1/GIP双受体激动剂DA3-CH可抑制大鼠癫痫持续状态导致的海马神经元凋亡,减少神经元丢失,具有神经保护作用。     

锂-匹罗卡品致痫大鼠脑组织中驱动蛋白家族成员17的表达情况

目的 观察驱动蛋白家族成员17 (KIF17)在锂-匹罗卡品致痫大鼠海马和颞叶皮质表达的变化,探讨其在癫痫发生、发展中的作用.方法 采用氯化锂-匹罗卡品诱导癫痫大鼠模型,将49只雄性正常Wistar大鼠采用随机数字表法分成实验组(n=42)和对照组(n=7),实验组又分为6个亚组(n=7):包括癫痫后24h、72 h、7d、14 d、1个月、2个月组.运用蛋白质印迹法检测KIF17在致痫大鼠海马和颞叶皮质的表达变化,免疫荧光双标染色法确定KIF17的表达部位.结果 大鼠海马KIF17蛋白表达在癫痫持续状态(SE)后开始增高[积分吸光度(L4)比值:24 h 0.516±0.196、72 h0.742±0.313],在癫痫后7d时达到高峰(0.888±0.319),之后逐渐降低(14 d 0.770±0.271、1个月0.742±0.261、2个月0.714±0.271),但均显著高于对照组(0.495±0.203),差异均有统计学意义(t=7.051、4.974、7.419、8.795、8.264、6.676,均P＜0.05).大鼠颞叶皮质KIF17蛋白的表达在SE后24h时开始持续增高,并在30 d时达到高峰,且IA比值均明显高于对照组.免疫荧光双标染色法显示KIF17蛋白主要存在于神经元,包括兴奋性神经元和抑制性神经元,而在星形胶质细胞中不表达.结论 KIF17在锂-匹罗卡品致癫痫模型的发生发展过程中可能起着重要的作用.     

氯化锂-匹罗卡品致痫幼鼠脑内c-jun和c-fos蛋白的表达

目的 探讨幼鼠癫痫持续状态(SE)后脑内c-jun和c-fos蛋白的表达。方法 采用氯化锂-匹罗卡品腹腔注射制成幼鼠SE模型。应用免疫组化及常规病理检查方法观察c-jun和c-fos蛋白的表达。结果 注射匹罗卡品1h后在海马结构和大脑皮质的大部分区域出现少量的c-jun和c-fos免疫反应阳性细胞，3～6h强烈表达，并达到高峰，24h后明显减弱，72h近乎正常对照水平。地西泮干预组幼鼠脑内有少量的c-jun和c-fos免疫反应阳性细胞，略多于生理盐水对照组。海马结构区的c-jun和c-fos免疫反应阳性细胞明显高于皮质区。SE组幼鼠的CA1、CA3和齿状回区可见到许多神经元发生变性和坏死。结论 c-jun和c-fos蛋白表达及其表达程度与SE后脑损伤的部位和分布有关；地西泮有对抗匹罗卡品致病作用，可能对脑组织有保护作用。     

普瑞巴林对慢性颞叶癫癎大鼠海马凋亡调控基因的影响

目的通过观察普瑞巴林对匹罗卡品慢性癫癎大鼠海马区Bcl-2和Bax表达的影响,探讨普瑞巴林治疗癫癎的药理学机制及对大鼠海马神经元的抗凋亡作用。方法采用氯化锂-匹罗卡品化学诱导方法建立慢性颞叶癫癎模型。经腹腔注射普瑞巴林40mg(/kg·d)连续治疗3周,免疫组织化学染色和Western blotting法检测不同处理组大鼠海马区Bcl-2和Bax表达变化。结果与生理盐水对照组比较,模型组大鼠海马区Bcl-2和Bax表达水平显著升高(均P=0.000);与模型组比较,普瑞巴林治疗组大鼠海马区Bcl-2表达水平升高、Bax表达水平降低,组间差异具有统计学意义(均P=0.000)。结论新型抗癫癎药物普瑞巴林可通过降低慢性颞叶癫癎大鼠海马区Bax表达、上调Bcl-2表达而抑制细胞凋亡,发挥神经元保护作用。     

经皮三叉神经慢性电刺激对慢性癫痫大鼠海马和皮层内VGLUT1表达的影响

目的观察经皮三叉神经慢性电刺激(trigeminal nerve stimulation,TNS)对匹罗卡品诱导的癫痫大鼠海马和皮层内囊泡型谷氨酸转运体-1(vesicular glutamate transporter 1,VGLUT1)表达的影响,探讨TNS治疗癫痫的可能机制。方法将达到癫痫持续状态(status epilepticus,SE)的实验大鼠随机分为模型(Pilo)组和经皮三叉神经慢性电刺激(TNS)组;正常对照组给予同剂量的生理盐水代替Pilo腹腔注射,随后对照组和Pilo组均给予为期一个月的经皮三叉神经假性电刺激(TNS)组给予为期一个月的真性电刺激。通过免疫荧光双标和Western blot分析海马和皮层内VGLUT1的表达。结果 TNS组和Pilo组海马、皮层内VGLUT1的表达均呈先升高后下降的趋势,约在72 h左右达到高峰。在24 h时,TNS组海马和皮层内VGLUT1的表达均明显高于Pilo组(P<0.01),而在以后各个时间点均低于Pilo组(P<0.05,P<0.01)。结论经皮三叉神经慢性电刺激能够减少癫痫大鼠脑内VGLUT1的表达,从而影响谷氨酸能神经元兴奋性神经递质的传递,推测TNS抗癫痫的机制可能与此有关。     

TGFβ1 Treatment Reduces Hippocampal Damage, Spontaneous Recurrent Seizures, and Learning Memory Deficits in Pilocarpine-Treated Rats

Studies have demonstrated the neuroprotective activity of transforming growth factor beta-1 (TGFβ1), protecting neurons against different kinds of insults. However, the role of exogenous TGFβ1 in the neuronal damage following status epilepticus (SE) and the related spontaneous recurrent seizures (SRS) is unknown. The present study aimed to determine the effect of intranasal TGFβ1 administration on SRS and cognitive function following lithium–pilocarpine-induced SE and associated hippocampal damage. We found that intranasal TGFβ1 significantly attenuated the hippocampal insults marked by hematoxylin and eosin, terminal deoxynucleotidyl transferase dUTP nick end labeling, and Fluoro-Jade B staining by 24, 48, and 72 h after SE was induced. The expression of the apoptosis-suppressing protein, Bcl-2, was elevated, whereas the expression of the apoptosis-promoting proteins, Bax and Caspase-3, was suppressed in TGFβ1-treated rats compared to rats without TGFβ1 treatment by 24, 48, and 72 h following induction of SE. The seizure number, severity, and duration of SRS over a 1-month period of monitoring starting 15 days after SE induction as well as the cognitive deficits detected 45 days after SE induction were significantly reduced in TGFβ1-treated rats compared to those without TGFβ1 treatment. Our results indicate that intranasal delivery of TGFβ1 immediately after SE induction not only protected against SRS but also improved cognitive function. The anti-epileptogenic properties of TGFβ1 may be related to its effect of neuroprotection or to its effect of apoptosis pathway changes.     

缺血性脑卒中患者外周血CBX7的表达水平变化及其通过Nrf2/HO-1通路对神经元凋亡的影响

目的 探讨Polycomb chromobox7(CBX7)对氧糖剥夺/再灌注(Oxygen-glucose deprivation/reperfusion,OGD/R)神经元的影响及其可能的作用机制.方法 收集缺血性脑卒中(Ischemic stroke,IS)组患者和体检健康人群(对照组)外周血,分离外周血单个核细...     

缺氧-复氧对大鼠海马培养神经元Bcl-2、Bax表达和神经元凋亡的影响

目的 观察缺氧-复氧对体外培养海马神经元Bcl-2和Bax表达和神经元凋亡的影响。方法 取培养12d的海马神经元，置于恒温(36℃)密闭容器内，连续充以无氧气体[90％(体积分数)N2、10％(体积分数)CO2]，在缺氧条件下继续培养4h后，再于常氧培养箱内复氧培养24h和72h。于不同时间观察神经元存活数，并分别用抗Bcl-2和Bax抗血清进行免疫组织化学染色，观察缺氧-复氧后大鼠海马培养神经元Bcl-2和Bax表达。并用原位末端标记(TUNEL)法和流式细胞术分别检测缺氧-复氧对体外培养海马神经元凋亡的影响。结果 缺氧-复氧后24～72h,海马神经元对Bcl-2的表达逐渐减弱，对Bax的表达逐渐增强，对Bax／Bcl-2比值逐渐增大，凋亡神经元百分率逐渐增多。结论 缺氧-复氧后24～72h神经元凋亡的发生与神经元Bcl-2表达逐渐减弱，Bax表达逐渐增强，Bax／Bcl-2比值逐渐增大有关。     

Mild hypothermia increases Bcl-2 protein expression following global cerebral ischemia.

Mild hypothermia protects the brain against experimental ischemia, but the reasons are not well known. We examined whether the protective effects of mild hypothermia could be correlated with alterations in expression of Bcl-2, an anti-apoptotic protein in a rat model of transient global ischemia. Following 10 min of forebrain ischemia, hippocampal neurons were examined 72 h later for survival, expression of Bcl-2 family proteins and apoptosis. Intraischemic mild hypothermia was applied for 3 h (33 degrees C, isch-33) or normal body temperature was maintained (37 degrees C, isch-37). Survival of CA1 neurons was significantly improved in the isch-33 group compared to the isch-37 group (90 vs. 53% survival; P<0.01). The proportion of Bcl-2-positive cells among surviving CA1 neurons in the isch-33 group was increased compared to that of sham and isch-37 groups (P<0.01). Bax expression in CA1 was no different between sham and isch-33 groups, but was significantly decreased in isch-37 (P<0.05). TUNEL staining was positive in many isch-37 CA1 neurons, but absent in isch-33. Utilizing electron microscopy, more cells meeting criteria for apoptosis were observed in the isch-37 than isch-33. These data suggest that mild hypothermia attenuates apoptotic death, and that this protection may be related to increases in Bcl-2.     

缺氧复氧大鼠海马星形胶质细胞凋亡及Bcl-2和Bax蛋白表达的时间进程变化及异丙酚干预效应

BACKGROUND： Cerebral hippocampal astrocytes are more sensitive.to ischemic injury than neurons. Hypoxic-ischemic brain injury induces profound astrocyte apoptosis, and propofol may protect against astrocyte apoptosis.
OBJECTIVE： To verify the protective effects of propofol against astrocyte apoptosis and to investigate anti-apoptotic Bcl-2 and pro-apoptotic Bax expression in primary cultures of rat hippocampal astrocytes exposed to hypoxia-reoxygenation for different periods of time following propofol treatment.
DESIGN, TIME, AND SETTING： In vitro neural immunocytochemistry was performed at the Central Laboratory of Yunyang Medical College between September 2007 and March 2008.
MATERIALS： A total of 30 Wistar rats, aged 1-3 days, wJth equal numbers of males and females, were included for isolation and culture of .hippocampal astrocytes.
METHODS： Hippocampal astrocytes were purified and cultured for 3 weeks and treated with four culture conditions： 50 μL Hank＇s solution （normal control）; 0.2 mL/L Intralipid; 50 μL Hank＇s solution for 10 minutes followed by hypoxic incubation for 4 hours and normoxic incubation for 12, 24, 36, 48, 60 or 72 hours; propofol （250 μmol/L final） for 10 minutes followed by hypoxic incubation for 4 hours and normoxic incubation for 12, 24, 36, 48, 60 and 72 hours.
MAIN OUTCOME MEASURES： （1） Morphologic changes in hippocampal astrocytes. （2） Levels of astrocyte apoptosis and Bcl-2 and Bax expression.
RESULTS： Hypoxia and reoxygenation increased apoptosis over time, with Bcl-2 expression peaking at 24 hours and decreasing gradually （P 〈 0.01 ）; Bax expression peaked at 72 hours （P 〈 0.01）; the ratio of Bcl-2/Bax was 1.4, 0.8, and 0.6, respectively, at 24, 48 and 72 hours. Non-apoptotic astrocytes showed significant proliferation and swelling. Propofol treatment decreased apoptosis after hypoxia-reoxygenation （P 〈 0.01）, as well as Bct-2 and Bax expression （P 〈 0.05, P 〈 0.01）, with Bcl-2/Bax ratios of 1.6-1.8. Propofol treatmentalso blocked astrocyte proliferation and swelling. No apoptotic cells or Bcl-2/Bax expression was detected in astrocytes cultured in Hank＇s or Intralipid solution.
CONCLUSION： Propofol protects astrocytes against injury caused by hypoxia and reoxygenation via a mechanism that involves maintaining high ratios of Bcl-2/Bax.     

3-硝基丙酸预处理对大鼠局灶性脑缺血Bcl-2和Bax mRNA表达的影响

目的探讨3-硝基丙酸(3-NPA)预处理对大鼠局灶性脑缺血半暗带Bc l-2和Bax mRNA表达的影响。方法将大鼠腹腔注射3-NPA 20 mg/kg,3 d后制作局灶性脑缺血再灌注模型;采用逆转录聚合酶链反应,观察3-NPA预处理对脑缺血再灌注1 h、6 h、12 h、24 h及48 h额顶部皮质Bc l-2和Bax mRNA表达的影响,并与假手术组和缺血再灌注组比较。结果与假手术组比较,缺血再灌注组和3-NPA预处理组各时间点Bc l-2和Bax mRNA表达极显著增强(均P<0.01);与缺血再灌注组比较,3-NPA预处理组各时间点Bc l-2mRNA表达显著增强(均P<0.05),再灌注12~48 h Bax mRNA的表达显著降低(均P<0.05)。结论增强Bc l-2的表达、抑制Bax的表达,可能是3-NPA预处理抑制细胞凋亡、诱导脑缺血耐受的机制之一。     

Pre-ischemia electro-acupuncture potentiates the expression of Bcl-2 and transforming growth factor-beta 1 in rat brains

The expression of the anti-apoptotic molecules Bcl-2 and transforming growth factor-beta 1 is known to confer protective effects on the cerebral ischemia-reperfusion injury.The current study investigated the expression levels of Bcl-2 and transforming growth factor-beta 1 in response to multiple pre-ischemia electro-acupuncture at acupoints Zusanli(ST36)and Fengchi(GB20) stimulation.Rats were divided into five groups:uninjured,control,non-acupoint,GB20 and ST36. Rats in the non-acupoint,GB20 and ST36 groups received 30 minutes(3 times or 18 times)of electro-acupuncture stimulation before experimental cerebral ischemia was induced.Bcl-2 and transforming growth factor-beta 1 were found to be significantly increased in the ST36 groups with either 3 or 18 electro-acupuncture treatments(P<0.05).The production was higher with 18 electro-acupuncture treatments in the ST36 groups(P<0.05).In the GB20 groups,significant increase was only observed in transforming growth factor-beta 1 with 18 electro-acupuncture treatments(P<0.05).No significant elevation of the level of transforming growth factor-beta 1 was observed in the non-acupoint groups.However,the production of Bcl-2 increased with 18 treatments in the non-acupoint groups(P<0.05).The data suggest that multiple pre-ischemia electro-acupuncture at ST36 was effective in conferring neuroprotective effect on the brain by means of upregulation of Bcl-2 and transforming growth factor-beta 1 and the effect was increase with the number of treatment.     

Expression of Bax and Bcl-2 protein in the gerbil hippocampus following transient forebrain ischemia and its modification by phencyclidine

Abstract

To determine the effect of phencyclidine (a noncompetitive NMDA receptor antagonist) on expression of Bax and Bcl-2 (apoptosis-regulating proteins) in gerbil hippocampus after transient forebrain ischemia, brain sections were immunohistochemically evaluated 48, 72, 96 hand 7 days following ischemia. In ischemic control animals, the expression of Bax in CA 7 neurons was increased with time and peaked at 72 h, then disappeared at 96 h. In the phencyclidine (5 mg kg^-1 , intraperitoneally)-treated animals, the intensity of Bax expression at 72 h was weaker than that of ischemic control animals. Furthermore, at 96 h, Bax expression was still observed in CA1 neurons. No expression of Bcl-2 in the CA1 neurons was detected in either control or phencyclidine-treated animals. From these results, it is possible that NMDA receptor antagonists exert their preventive effect against delayed neuronal death through inhibition of Bax protein expression, although the precise relationship between the function of Bax protein and delayed neuronal death is still unclear. [Neural Res 1997; 19: 629-633]     

巴曲酶对沙土鼠海马CA1区神经元保护作用及其机制的研究

目的探讨巴曲酶对沙土鼠前脑缺血再灌注神经元保护作用的可能机制及时效关系。方法实验动物随机分为10组,分别为缺血再灌注后0h、1h、3h、6h、12h、24h、48h和72h组、假手术组及对照组,各个时间点分别给予腹腔注射巴曲酶8BU／kg,采用免疫组化SP法检测海马CA1区的Bcl-2、eNOS、VEGF及Akt蛋白的表达,光镜下计算沙土鼠海马CA1区的神经元阳性细胞数。并行电镜下神经元超微结构的观察。结果使用巴曲酶后,Bcl- 2、eNOS、VEGF及Akt蛋白表达明显增加,神经元阳性细胞数在巴曲酶各时间点与对照组比较差异有显著性(P< 0．01),而巴曲酶0h-24h时间点之间比较差异无统汁学意义(P>0．05);Bcl-2、eNOS及VEGF神经元阳性细胞数在0h-24h时间点与48h-72h时间点比较差异有统计学意义(P<0．05);超微结构显示0h-24h时间点较48h及 72h时间点抗细胞凋亡明显。结论巴曲酶脑保护作用的机制可能是通过上调Bcl-2、eNOS、VEGF及Akt蛋白的表达;72h内使用巴曲酶具有神经元保护作用,但在0h-24h时间点使用较48h及72h时间点保护作用更为明显。