当归对缺氧缺血性脑损伤幼年大鼠神经元的保护作用

目的探讨脑缺氧缺血对幼年大鼠海马齿状回神经元的影响及当归注射对其保护作用。方法取7日龄健康SD新生大鼠33只,随机分为对照组、缺氧组和当归组各11只。缺氧组和当归组新生大鼠在无菌环境下结扎左侧颈总动脉,术后护理2 h后置于三气培养箱持续缺氧2 h/d,连续7 d,制作新生鼠缺氧缺血性脑损伤模型,对照组仅行假手术,不结扎左侧颈总动脉、不缺氧。术后第8 d开始,缺氧组和对照组大鼠经腹腔注射生理盐水（8 ml/Kg）,连续7 d;当归组用等量当归注射液（250 g/L）代替生理盐水。于生后第40 d取大鼠脑组织,常规石蜡包埋、经海马切片,行神经元特异性烯醇化酶（NSE）免疫组化染色,图像分析海马齿状回NSE阳性细胞的积分光密度（IOD）值。结果缺氧组大鼠海马齿状回NSE阳性细胞的IOD值较对照组降低,而当归组NSE阳性细胞的IOD值较缺氧组增高。结论脑缺氧缺血可降低幼年大鼠海马齿状回NSE的表达,而当归注射液对缺氧缺血性脑损伤幼年大鼠神经元可能具有保护作用。     

川芎嗪对内毒素性休克大鼠脑损伤的作用

目的:研究川芎嗪(ligustrazine,Lig)对内毒素性休克大鼠脑损伤的作用并探讨其作用机制。方法:将48只健康Wistar大鼠随机分为正常组、LPS组和LPS+Lig组,每组16只,上述每组再分为2个亚组:6 h组和12 h组,各8只大鼠。以尾静脉注射5 mg/kg脂多糖(LPS)建立内毒素性休克大鼠模型,腹腔注射盐酸川芎嗪注射液200 mg/kg,分别在相应时点进行眼球摘除采血并断头取脑组织,ELISA法检测血清中神经元特异性烯醇化酶(neuron-specific enolase,NSE)含量,脑组织匀浆检测一氧化氮(nitric oxide,NO)含量,TUNEL染色法检测脑组织中海马区细胞凋亡情况,Western blot法检测Bax和Bcl-2蛋白的表达。结果:与LPS组相比,川芎嗪能降低内毒素性休克所致大鼠NSE和NO含量的上升,同时抑制海马区细胞的凋亡,增加Bax的蛋白表达并降低Bcl-2的蛋白表达。结论:川芎嗪通过降低NSE和NO含量,减轻内毒素性休克大鼠的脑损伤程度,可能与其调节Bax和Bcl-2蛋白的表达有关。     

大鼠创伤性脑损伤后细胞凋亡及NOS阳性细胞的变化

目的：探讨大鼠创伤性脑损伤后不同时相皮质、海马、隔区细胞凋亡及NOS、ChAT阳性细胞的变化。方法：采用大鼠自由落体脑损伤模型，伤后1、2、3、4、5、7、10d取脑切片，经Nissl染色，用TUNEL法检测细胞凋亡，NADPH—d组化染色观察NOS阳性细胞，ChAT免疫组化染色观察隔区ChAT阳性细胞。结果：Nissl染色可见损伤侧海马CA2、CA3区锥体细胞层细胞消失或紊乱。损伤区周围皮质凋亡细胞伤后3d达到高峰；损伤侧海马凋亡细胞伤后5d达到高峰；损伤侧隔区凋亡细胞7d达到高峰。正常侧上述脑区各时相点均未见到凋亡细胞。损伤区周围皮质、损伤侧海马和隔区iNOS阳性细胞数量明显增加。损伤侧隔区ChAT阳性神经元也明显减少。结论：大鼠创伤性脑损伤后损伤区周围皮质和损伤侧海马、隔区细胞凋亡数量的变化与伤后时程有关。伤后细胞iNOS表达增加是导致细胞凋亡的因素。     

VEGF转基因神经干细胞移植对大鼠急性放射性脑损伤后脑组织中NSE表达的影响

目的： 探讨血管内皮生长因子（VEGF）转染的神经干细胞(NSC)对放射性脑损伤的影响。方法：将Sprague-Dawley(SD)大鼠随机分为对照组、放射性脑损伤组、NSC和转染VEGF的NSC治疗组, 放射性脑病模型成功制备后1周脑内移植转染VEGF的NSC,移植后1周、2周、3周、4周和6周末取其脑组织,用免疫组织化学法测定神经元特异性烯醇化酶(NSE)阳性细胞数,Western blotting法检测NSE蛋白表达情况。结果：与对照组相比, 放射性脑损伤组大鼠脑组织中NSE阳性细胞数显著下降(P<0.05);与放射性脑损伤组相比,NSC治疗组和转染VEGF的NSC治疗组在移植3周后NSE阳性细胞数明显增加(P<0.05), 与NSC治疗组比较,在移植6周末转染VEGF的NSC治疗组NSE的阳性细胞数增加更明显(P<0.05)。与放射性脑损伤组比,NSC治疗组和转染VEGF的NSC治疗组在移植3周末NSE蛋白表达量明显增加(P<0.05), 与NSC治疗组比较,在移植6周末转染VEGF的NSC组NSE蛋白表达量增加更明显(P<0.05)。结论：转染VEGF的NSC移植可提高放射性脑损伤组织中NSE的含量。     

大鼠创伤性脑损伤后星形胶质细胞的变化

目的：探讨大鼠创伤性脑损伤后星形胶质细胞的形态学变化及GFAP和NOS的表达情况。方法：采用大鼠自由落体脑损伤模型，伤后1、3、7d取脑切片，行Nissl染色以及GFAP免疫组化和NADPH—d组化单标记及双标记染色。结果：损伤区周围皮质GFAP阳性细胞胞体增大、突起增粗增长，GFAP阳性细胞数量与正常侧及对照组相比，伤后1d即有明显增加，伤后3d、7d数量持续增加；损伤侧海马CAI～3区和DG各层GFAP阳性细胞排列紊乱，胞体增大、突起增粗增长，GFAP阳性细胞数量与正常侧及对照组相比则无明显变化。损伤区周围皮质、损伤侧海马NOS阳性细胞数量明显增加。伤后3d损伤区周围皮质和损伤侧海马中GFAP与NOS双标细胞分别占GFAP阳性细胞的14．2％和13．4％左右。结论：大鼠创伤性脑损伤后大量的星形胶质细胞活化、GFAP表达增加并且部分转化为NOS阳性细胞，提示其参与了脑组织的损伤与修复过程。     

癫痫患儿血清神经元特异性烯醇化酶水平变化及临床意义

目的　研究癫痫患儿癫痫发作后血清神经元特异性烯醇化酶 (NSE)的变化。方法　采用放射免疫分析法对11例癫痫患儿发作后 4 8h内血清中NSE进行测定 ,并与 2 2例正常组进行对照。结果　 2 2例正常对照组血清中NSE进行测定值为 (6 .4 6± 3.35 ) μg/L ,癫痫发作组血清中NSE为 (17.4 5± 10 .1) μg/L ,两组比较有显著性差异 (P≤ 0 .0 1)。结论　癫痫发作组NSE水平明显高于正常对照组 ,提示癫痫发作后有一定程度的脑神经元损害。     

儿童早期脑损伤血清神经元特异性烯醇化酶与发育行为异常的相关性研究

目的 探讨儿童早期脑损伤血清神经元特异性烯醇化酶(neuronspecific enolase,NSE)与后期发育行为异常的相关性.方法 选择我院出生的重度、中度、轻度HIE患儿各15例以及15例健康儿,采用酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)测定血清中神经元特异性烯醇化酶的浓度,并进行新生儿行为神经测定(neonatal behavioral neurological assessment,NBNA)评分以及ASQ发育筛查系统(ASQ developmental screening system,简称ASQ-C评价).结果 两组儿童在性别、体重、分娩方式等指标上均不存在显著差异(P>0.05);重度、中度、以及轻度新生儿缺氧缺血脑病(hypoxic-ischemic encephalopathy,HIE)患儿之间血清NSE值存在显著差异(P<0.05),血清中NSE值随临床病征的加重而升高;实验组中重度、中度、以及轻度HIE患儿的血清NSE值均高于对照组,且差异存在统计学意义(P<0.05);重度、中度、以及轻度HIE患儿之间NBNA评分值存在显著差异(P<0.05),NBNA评分值随临床病征的加重而降低;实验组中重度、中度、以及轻度HIE患儿的NBNA评分值均低于对照组,且差异存在统计学意义(P<0.05);不同NBNA评分组之间血清NSE值存在显著差异(P<0.05),血清NSE值随着NBNA评分的增加而降低;实验组与对照组比较,重度、中度和轻度HIE组沟通区异常、大运动能区异常、精细动作能区异常较对照组皆存在显著差异(P<0.05);解决能力区异常、个人-社会能区异常上实验组与对照组不存在显著差异(P>0.05).结论 NSE以及NBNA评分与儿童早期脑损伤患儿的严重程度以及后期的发育行为异常相关.     

急性创伤性脑损伤患者初期血清NSE含量及判断限值的探讨

目的：探讨血清NSE的含量在脑损伤的诊断和评估伤情及预后早期判断中的临床意义,建立其判断限值。方法：采用发光免疫法,分析了79例急性脑损伤患者初期血清NSE的含量,并与98例正常人组及86例非神经系统疾病患者的检测结果作比较。对不同程度、不同预后的脑损伤患者血清NSE含量的差别,进行显著性检验,并对GCS评分与NSE间的关系作相关分析,通过对不同判断值的临床可用性能评价确定脑损伤患者血清NSE的判断限值。结果：急性创伤性脑损伤组血清NSE含量显著高于正常人组及非神经系统疾病组（P〈0.001）,急性重型脑损伤组患者血清NSE的浓度〉中型组〉轻型组（P〈0.01,〈0.05）,相关分析显示,急性脑损伤患者初期血清NSE的浓度与GCS评分呈负相关（r=-0.8921,P〈0.01）。并发现脑损伤初期血清NSE浓度〈30ng/ml组和（30～60）ng/ml组及≥60ng/ml组的急性创伤性脑损伤患者的预后良好率分别为97.4%和75%及18.8%,残废率分别为2.6%和20.8%及50.0%,死亡及植物生存率分别为0%和2.6%及31.2%,三组间预后的构成比有显著性差异（P〈0.05,〈0.01）。临床可用性能评价结果显示,判断值提高至22.0ng/ml时,准确度和可用度均名列第一,其诊断试验的特异性和敏感性分别达88.4%和62%。结论：血清NSE含量测定不仅是脑损伤诊断和伤情评估的一项量化指标,而且有望成为早期判断预后的手段之一。脑外伤后血清NSE浓度≥22ng/ml应首先考虑脑损伤的存在。     

创伤性脑损伤后大鼠皮质AQP4表达的变化

目的 观察创伤性脑损伤后大鼠皮质水通道蛋白4(aquaporin4,AQP4)表达的变化.方法 采用Feency法建立脑损伤大鼠模型,取成年健康雄性wistar大鼠60只,随机分为6组,So、S1、S4、S1W、S2W及正常对照组.采用免疫组化和免疫印迹方法进行各组AQP4表达变化的观察及检测,进行图像分析和统计学处理...     

金纳多对体外循环脑损伤的保护作用

目的观察金纳多对体外循环脑损伤的保护作用。方法择期行风湿性心脏病二尖瓣置换术患者40例，美国麻醉医师协会分级Ⅱ～Ⅲ级，按随机数字表法随机分为银杏叶提取物组（EGB组）和对照组各20例。银杏叶提取物组（20例）：体外循环管道预充乳酸林格氏液中加入银杏叶提取物注射液1mg／kg；对照组（20例）：体外循环预充乳酸林格氏液。分别于体外循环开始前（T1）、体外循环开始后30min（T2）、体外循环结束时（L）、体外循环结束后2h（T4）、体外循环结束后6h（T5）、体外循环结束后12h（T6）、体外循环结束后24h（T7）7个时间点同步采集桡动脉及颈内静脉血，动脉血进行血气分析，静脉血测定血清S-100β蛋白、神经元特异性烯醇化酶（neuron specific enolase，NSE）浓度。结果CPB开始后即有血清S-100β蛋白和NSE浓度升高，EGB组S-100β蛋白和NSE浓度明显低于对照组，且其达峰时间提前。结论银杏叶提取物能改善脑氧供需平衡，明显减少血清脑损伤标志物S-100β蛋白和NSE的浓度升高，有明确的脑保护作用。     

Neuroprotective effects of resveratrol against traumatic brain injury in immature rats

Childhood trauma resulting in traumatic brain injury (TBI) due to accidents and abuse is the major cause of death and dysfunction in the young. Since there are no approved specific pharmacological agents that block the progression of the secondary injury, the current management of TBI is mainly supportive. We aimed to determine the effect of resveratrol on hippocampal damage and behavioral deficits in 7-day-old rat pups subjected to contusion injury. Resveratrol was injected intraperitoneally at the doses of 100 mg/kg of body weight immediately after induction of traumatic injury. Hippocampal damage was examined by cresyl violet staining and behavioral alterations were evaluated using open field and novel object recognition tests 2 weeks after trauma. Histopathological evaluation showed that treatment with a single dose of 100 mg/kg resveratrol (i.p.) after the trauma significantly ameliorated the trauma induced hippocampal neuron loss at ipsilateral and contralateral hippocampal brain regions of rats. Additionally, treatment with resveratrol decreased anxiety and increased cortex/hippocampus dependent memory of animals subjected to blunt head trauma. These results show that acute treatment of resveratrol has a neuroprotective role against trauma induced hippocampal neuron loss and associated cognitive impairment in rats.     

Sex differences in XIAP cleavage after traumatic brain injury in the rat

Sex influences histological and behavioral outcomes following traumatic brain injury (TBI), but the underlying sex-dependent pathomechanisms regulating outcome measures remain poorly defined. Here, we investigated the TBI-induced regulation of the X-linked inhibitor of apoptosis protein (XIAP) that, in addition to suppressing cell death by inhibition of caspases, is involved in signaling cascades, including immune regulation and cell migration. Since estrogen has been shown to have anti-apoptotic properties, we specifically examined sex differences and the influence of estrogen on XIAP processing after TBI. Sprague–Dawley male (TBI-M), female (TBI-F), ovariectomized female (TBI-OVX) and ovariectomized females supplemented with estrogen (TBI-OVX + EST) were subjected to moderate (1.7–2.2 atm) fluid percussion (FP) injury. Animals were sacrificed 24 h after FP injury; cortical tissue (ipsilateral and contralateral) was dissected and analyzed for XIAP processing by immunoblot analysis (n = 6–7/group) or confocal microscopy (n = 2–3/group). Significant differences in XIAP cleavage products in the ipsilateral cortex were found between groups (p < 0.03). Post hoc analysis showed an increase in XIAP processing in both TBI-F and TBI-OVX + EST compared to TBI-M and TBI-OVX (p < 0.05), indicating that more XIAP is cleaved following injury in intact females and TBI-OVX + EST than in TBI-M and TBI-OVX groups. Co-localization of XIAP within neurons also demonstrated sex-dependent changes. Based on these data, it appears that the processing of XIAP after injury is different between males and females and may be influenced by exogenous estrogen treatment.     

脑外伤小鼠海马水通道蛋白-9表达的变化

目的探讨脑外伤小鼠海马水通道蛋白-9(aquaporin 9,AQP9)表达的变化。方法40只Balb/c小鼠随机分为假手术组(10只)和脑外伤组(30只)。脑外伤组依据脑外伤的不同时间点再分6 h、1 d、3 d三个小组,每组10只。免疫组化和Western blot检测各组小鼠海马AQP9蛋白的表达。RT-PCR方法检测各组小鼠海马AQP9 mRNA的表达变化。结果免疫组化和Western blot结果发现,脑外伤组小鼠海马AQP9蛋白表达量明显高于假手术组(P<0.05);RT-PCR结果也发现,脑外伤组小鼠海马AQP9 mRNA表达量明显高于假手术组(P<0.05)。结论脑外伤小鼠海马AQP9表达明显升高。     

Chelation of neurotoxic zinc levels does not improve neurobehavioral outcome after traumatic brain injury

Increases of synaptically released zinc and intracellular accumulation of zinc in hippocampal neurons after traumatic or ischemic brain injury is neurotoxic and chelation of zinc has been shown to reduce neurodegeneration. Although our previous studies showed that zinc chelation in traumatically brain-injured rats correlated with an increase in whole-brain expression of several neuroprotective genes and reduced numbers of apoptotic neurons, the effect on functional outcome has not been determined, and the question of whether this treatment may actually be clinically relevant has not been answered. In the present study, we show that treatment of TBI rats with the zinc chelator calcium EDTA reduces the numbers of injured, Fluoro-Jade-positive neurons in the rat hippocampus 24 h after injury but does not improve neurobehavioral outcome (spatial memory deficits) 2 weeks post-injury. Our data suggest that zinc chelation, despite providing short-term histological neuroprotection, fails to improve long-term functional outcome, perhaps because long-term disruptions in homeostatic levels of zinc adversely influence hippocampus-dependent spatial memory.     

Glutamine decreases intestinal nuclear factor kappa B activity and pro-inflammatory cytokine expression after traumatic brain injury in rats

Objective: To investigate whether glutamine supplementation modulates intestinal nuclear factor kappa B (NF-κB) activity and pro-inflammatory cytokine expression after traumatic brain injury (TBI) in rats. Materials and methods: Right parietal cortical contusion in male rats was made by the weight-dropping method. After trauma, the rats were randomly given chow alone or glutamine mixed chow for 5 d. Gut samples were extracted at 5 d postinjury. We measured NF-κB binding activity by electrophoretic mobility shift assay; NF-κB subunits p50 and p65 expression by immunohistochemistry; the concentrations of interleukin-1β, tumor necrosis factor-α and interleukin-6 by enzyme-linked immunosorbent assay; intestinal mucosal morphological changes by histopathological study and electron microscopy; and apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. Results: Administration of glutamine following TBI could decrease NF-κB binding activity, NF-κB p65 protein expression and concentrations of pro-inflammatory cytokines in the gut. TBI-induced damage of gut structure was ameliorated after glutamine supplementation. Conclusion: The results of the present study suggest that the therapeutic benefit of post-TBI glutamine supplementation might be due to its inhibitory effects on intestinal NF-κB activation and pro-inflammatory cytokine expression. Received 14 May 2007; returned for revision 9 July 2007; accepted by I. Ahnfeld-R?nne 16 August 2007     

Cell-specific DNA fragmentation may be attenuated by a survivin-dependent mechanism after traumatic brain injury in rats

Survivin attenuates apoptosis by inhibiting cleavage of some cell proteins by activated caspase-3. We recently discovered strong up-regulation of survivin, primarily in astrocytes and a sub-set of neurons, after traumatic brain injury (TBI) in rats. In this study we characterized co-expression of survivin with activated caspase-3 and downstream DNA fragmentation (TUNEL) in astrocytes and neurons after TBI. Western blot analysis revealed significant time-dependent increases in active caspase-3 between 5 and 14 days post-injury. No difference was observed between the proportion of survivin-positive and survivin-negative cells labeled with active caspase-3 at 5 or 7 days post-injury, as indicated by dual fluorescent immunostaining. Labeling of survivin-negative cells with TUNEL was, however, significantly greater than for survivin-positive cells, suggesting that expression of survivin may attenuate DNA cleavage and progression of apoptosis. A higher proportion of astrocytes than neurons accumulated active caspase-3. In contrast, co-localization with TUNEL was significantly higher for neurons than for astrocytes. These data suggest that survivin expression may attenuate DNA cleavage and cell death, and that this mechanism operates in a cell type-specific manner after TBI.     

Memory and executive functions correlates of self-awareness in traumatic brain injury

Objective: The purpose of this study was to investigate the contribution of executive functions (EF) components and episodic and working memory variables, as well as clinical and demographic factors, to awareness of cognitive ability in traumatic brain injury (TBI).

Methods: Sixty-five TBI patients (mild: n?=?26; moderate/severe: n?=?39) took part in the study. Independent stepwise regression models were calculated for EF and memory predictors, with awareness being measured by patient/informant discrepancy in the Patient Competency Rating Scale.

Results: Models with EF variables indicated that semantic verbal fluency and age are the best predictors of awareness, whereas models including mnemonic functions suggested verbal delayed episodic recall and TBI severity as predictors.

Conclusions: These results are discussed in relation to clinical implications, such as the need to focus efforts of rehabilitation in the cognitive abilities related to awareness, and theoretical models.     

Performance discrepancies on the California Verbal Learning Test: Second Edition (CVLT-II) after traumatic brain injury.

One hundred fourteen patients with traumatic brain injury (TBI), selected from a 5-year series of consecutive rehabilitation referrals, completed the California Verbal Learning Test -- Second Edition (CVLT-II) within 1 year after injury. Various performance contrasts (i.e., proactive interference, retroactive interference, rapid forgetting, and retrieval problems) were evaluated. Initial analyses revealed higher rates of rapid forgetting in the TBI group as compared to the standardization sample. Follow-up analyses between those patients with and without unusual degrees of rapid forgetting did not reveal any significant differences between these groups on demographic or neurological variables (p>0.10 for all variables). It is concluded that performance discrepancies on the CVLT-II should never be used in isolation to determine the presence or absence of acquired cerebral or memory impairment. However, regardless of the cause, such discrepancies may still be relevant for clinical treatment recommendations.