Neuroprotective effect of high-dose hyperbaric oxygenation on rats with acute cerebral infarction in super-early stage

BACKGROUND: Previously, only single short-time low-dose hyperbaric oxygenation (HBO) protocol was administrated to treat acute ischemic stroke in early stage and the conflicting results were obtained. There are few studies to report the outcome of administering long-time (can cover all the natural pathologic progression period) high-dose HBO to treat the disease. OBJECTIVE: To evaluate the therapeutic effect between two kinds of high-dose hyperbaric oxygenation on super-early stage of acute permanent middle cerebral artery occlusion (MCAO) in rats. DESIGN: A randomized controlled experimental study. SETTING: Beijing Tiantan Hospital, Capital Medical University; Beijing Research Institute of Neurosurgery. MATERIALS: Seventy-four male SD rats, aged 2.5 months old, weighing （280±20）g, were provided by the Animal Institute, Chinese Academy of Medical Sciences. Hyperbaric oxygenation device was hyperbaric air cabin in which there was a self-made pure oxygen animal experimental cabin (made in China). METHODS: This experiment was carried out in the municipal laboratory of Beijing Tiantan Hospital affiliated to Capital Medical University and Beijing Research Institute of Neurosurgery. ① Experimental intervention: All the rats were developed into models of permanent MCAO by suture embolism. Then, they were randomly divided into two HBO groups (9 hours and 18 hours) and control group, with 24 rats in each as well as 3-hour ultrastructure control group, with 2 rats. After being modeled for 3 hours, rats in the two HBO groups stayed in the hyperbaric cabin for 9 hours and 18 hours, separately. Rats in the 9-hour HBO group inhaled pure oxygen at hours 1, 3, 5, 7 and 9, and hyperbaric air at hours 2, 4, 6 and 8. Rats in the 18-hour HBO group inhaled pure oxygen at hours 1, 3, 5, 7, 9, 11, 13, 15 and 17, and hyperbaric air at hours 2, 4, 6, 8, 10 12, 14, 16 and 18. After being created into models, rats in the control group and 3-hour ultrastructure control group breathed room air. ② Experimental evaluation: Neurologic functions of rat models in the 9-hour and 18-hour HBO groups as well as control group were scored by Bederson and Garica two neurological grading systems at hours 14 and 28 and on day 5; Infarct volume of rat models in the two HBO groups and control group was measured at hour 24 and on day 5 with NIH image processing software Image J; The pathological changes of brain tissue in the brain infarct region and its opposite region of rat models in the two HBO groups and 3-hour ultrastructure control group were observed with a Philips EM 208S transmission electron microscope. MAIN OUTCOME MEASURES: ① Neurobehavioral outcome. ② Rat brain infarct volume. ③ Ultrastructure of brain tissue in the ischemic penumbra of infarct models at the different time points RESULTS: ① Neurobehavioral outcome: After treatment, Garica score in the 9-hour and 18-hour HBO groups was significantly higher than that in the control group (P < 0.01). Bederson score on day 5 after modeling in the 9-hour and 18-hour HBO groups was significantly lower than that in the control group (P < 0.01). ② Cerebral infarct volume: Cerebral infarct volume in the 9-hour and 18-hour HBO groups was significantly smaller than that in the control group at hour 24 and on day 5 after modeling (P < 0.01). In the 18-hour HBO group, infarct volume on day 5 after modeling was significantly larger than that at hour 24 after modeling (P < 0.05). ③In the 3-hour ultrastructure control group, astrocyte edema and neuron damage around the capillary in the infarct cerebral tissue significantly relieved in the rats which were subjected to HBO. CONCLUSION: High dose of HBO is highly efficient in reducing infarct volume and improving neurobehavioral outcome of rats with acute cerebral infarction, and also has an important role in inhibiting the pathological progression of ischemic brain tissue after cerebral infarction.     

脑血疏口服液对大鼠脑缺血再灌注损伤后血脑屏障的影响

目的 探讨脑血疏口服液对大鼠脑缺血再灌注损伤后血脑屏障的影响。方法 将120只SD大鼠随机分为3组:假手术组、对照组,脑血疏组; 采用线栓法建立大鼠左侧大脑中动脉闭塞再灌注模型,缺血2 h后拔出线栓,恢复灌注24 h; 采用Longa FZ 5级评分法进行大鼠神经功能缺损评分; TTC染色计算脑梗死体积百分比; 运用干-湿重法测脑含水率; 通过伊文思蓝( EB)含量反映血脑屏障的损伤程度; 免疫组化检测基质金属蛋白酶-9(MMP-9)的表达水平。结果(1)假手术组大鼠在神经功能缺损评分、脑梗死体积、脑含水率均低于对照组(P<0.01); 脑组织中EB含量和MMP-9表达水平较对照组低(P<0.01);(2)脑血疏组大鼠的神经功能缺损评分较低、脑梗死体积较小,脑水肿程度较轻; EB含量和MMP-9表达水平均较对照组明显减少(P<0.01)。结论 脑血疏口服液对大鼠脑缺血再灌注损伤后血脑屏障具有保护作用,其机制可能是通过抑制MMP-9的表达。     

针刺对急性脑挫裂伤大鼠脑组织内p-CREB表达的影响

目的探讨针刺对急性脑挫裂伤大鼠磷酸化的环磷酸腺苷反应元件结合蛋白(p-CREB)表达的影响。方法 48只雄性SD大鼠随机分为针刺治疗组、模型对照组、正常对照组,每组16只。模型对照组仅制备急性脑挫裂伤模型,针刺治疗组在建立模型后行针刺治疗,正常对照组不致伤。损伤后48 h、6 d取损伤或对应局部脑组织,采用免疫组化法观察p-CREB的表达变化,并进行统计学分析。结果伤后48 h、6 d,模型对照组损伤脑组织中p-CREB的表达较正常对照组升高(均P<0.05),针刺治疗组损伤脑组织中p-CREB表达较模型对照组显著升高(均P<0.05)。伤后6 d针刺治疗组p-CREB表达较伤后48 h降低(P<0.05)。结论针刺治疗对大鼠颅脑损伤后p-CREB的表达具有明显的促进作用,并呈一定规律性,提示p-CREB与颅脑损伤后脑组织的修复相关。     

二苯乙烯苷对脑缺血再灌注大鼠TrkA及Bcl-2表达的影响

目的探讨二苯乙烯苷（TSG）对脑缺血再灌注大鼠的神经保护作用及机制。方法将250～350g健康雄性sD大鼠共4组：对照组、模型组、小剂量TSG组和大剂量TSG组,每组24只。Longa线栓法制备大脑中动脉栓塞模型（MCAO）,按Longa的5级标准评分法评价神经功能缺损。再灌注后6h、24h、48h和7d共4个时间点处死大鼠。采用原位末端脱氧核苷酸转移酶标记法（TUNEL）检测神经细胞凋亡;采用原位杂交方法、免疫组化法检测TrkA、Bcl-2基因／蛋白表达变化。结果神经功能缺损评分显示造模各组各时间点均有明显的神经功能缺损症状,除6h时间点外,两个剂量TSG治疗组其余各时间点神经功能缺损评分明显低于模型组,差异有统计学意义（P〈0．05）;与模型组比较,两个剂量TSG组各时间点凋亡细胞减少,TrkA、Bcl-2基因／蛋白的表达明显上调,差异都有统计学意义（P〈0．05）。结论TSG可能通过增强脑缺血再灌注损伤后TrkA／Bcl-2通路的活性,起到神经保护作用。     

Acupuncture promotes functional recovery after cerebral hemorrhage by upregulating neurotrophic factor expression

Acupuncture is widely used in the treatment of cerebral hemorrhage,and it improves outcomes in experimental animal models and patients.However,the mechanisms underlying the effectiveness of acupuncture treatment for cerebral hemorrhage are still unclear.In this study,a model of intracerebral hemorrhage was produced by injecting 50μL autologous blood into the caudate nucleus in Wistar rats.Acupuncture at Baihui(DU20)and Qubin(GB7)acupoints was performed at a depth of 1.0 inch,12 hours after blood injection,once every 24 hours.The needle was rotated at 200 r/min for 5 minutes,For each 30-minute session,needling at 200 r/min was performed for three sessions,each lasting 5 minutes.For the positive control group,at 6 hours,and 1,2,3 and 7 days after induction of hemorrhage,the rats were intraperitoneally injected with 1 mL aniracetam(0.75 mg/mL),three times a day.The Bederson behavioral test was used to assess palsy in the contralateral limbs.Western blot assay was used to examine the expression levels of Nestin and basic fibroblast growth factor in the basal ganglia.Immunohistochemistry was performed to count the number of Nestin-and glial cell line-derived neurotrophic factor-positive cells in the basal ganglia.Acupuncture effectively reduced hemorrhage and brain edema,elevated the expression levels of Nestin and basic fibroblast growth factor in the basal ganglia,and increased the number of Nestin-and glial cell line-derived neurotrophic factor-positive cells in the basal ganglia.Together,these findings suggest that acupuncture promotes functional recovery after cerebral hemorrhage by increasing the expression of neurotrophic factors.The study was approved by the Committee for Experimental Animals of Heilongjiang Medical Laboratory Animal Center(approval No.2017061001)on June 10,2017.     

脑心通对缺血性大鼠脑组织核转录因子-κB、基质金属蛋白酶-9和肿瘤坏死因子-α的影响

目的探讨脑心通对缺血性大鼠脑组织核转录因子-κB(NF-κB)、基质金属蛋白酶-9(MMP-9)和TNF-α的影响。方法将110只健康雄性SD大鼠随机分为假手术组(30只),模型对照组(40只),脑心通治疗组(40只),每组再分为缺血再灌注后12 h、1 d、3 d、5 d、7 d五个时间点进行处理。采用Zea Longa法制备大脑中动脉闭塞缺血再灌注模型,利用MRI检查和Bederson评分筛选成功模型;每组五个时间点分别选取6只大鼠行MRI检查,动态观察使用脑心通胶囊后缺血脑组织变化过程,采用western blot、实时定量PCR分别检测梗死区脑组织NF-κB、MMP-9和TNF-α的蛋白和mRNA表达水平。结果脑心通治疗组脑梗死区域体积较模型对照组减少。脑心通治疗组NF-κB、MMP-9、TNF-α蛋白及其mRNA水平均明显低于模型对照组(均P0.05);假手术组MMP-9、TNF-α和NF-κB蛋白及其mRNA水平与脑心通治疗组差异无统计学意义。结论脑心通通过改变脑梗死区周围炎症因子表达水平来降低炎症反应,发挥神经保护作用。     

大鼠脑出血早期脑组织神经元型一氧化氮合酶mRNA表达及中药抵当汤干预对其的影响

目的 探讨大鼠脑出血（ICH）早期脑组织神经元型一氧化氮合酶（nNOS）mRNA表达,以及中药抵当汤干预对其的影响。方法 将72只大鼠随机分为ICH组、抵当汤组、NOS抑制剂组和对照组,采用立体定向技术注入自体不凝血建立实验性ICH模型,对照组注入等量的生理盐水;术后6h、24h及72h用Bederson3级标准法对大鼠进行神经功能障碍评定;然后断头取脑,以原位杂交方法检测脑组织中nNOS mRNA阳性细胞的表达。结果 神经功能缺损程度评分术后72h时ICH组与抵当汤组均见明显改善（均P〈0.05）,但抵当汤组的改善明显优于ICH组（P〈0．05）;ICH后6h血肿侧大脑皮质nNOS mRNA阳性细胞的表达较对照组已有明显增高,24h达高峰,72h时下降,但仍明显高于对照组（均P〈0．05）;抵当汤组血肿侧大脑皮质nNOS mRNA阳性细胞表达在各时间点较ICH组明显减少（均P〈0．05）,且与NOS抑制剂组变化规律一致。结论 抵当汤具有与NOS抑制剂相似的作用,通过抑制ICH后脑组织中nNOS mRNA的表达而产生脑保护作用。     

不同途径移植人脐血单核细胞对脑出血大鼠神经功能的影响

目的 观察经不同途径移植人脐血单核细胞14天后脑出血大鼠神经功能评分的变化,探讨干细胞移植的最佳途径。方法 采用自体血二次注血/退针法制作脑出血模型,经计算机断层扫描（computed tomography,CT）检查证实脑出血模型构建成功后,将从人脐血中分离出的新鲜人脐血单核细胞（human umbilical cord blood mononuclear cells,HUCBMC）分别经Wistar大鼠尾静脉、左心室及脑出血局部移植入实验动物体内,对照组造模成功后,不予治疗,自然转归。各组模型均于移植1、3、7、14天采用Longa评分法评价大鼠神经功能。结果 经尾静脉、左心室及脑出血局部移植HUCBMC的大鼠神经功能评分,经统计学处理后提示不同时间点大鼠的神经功能评分差异有显著性（F =131.87,P＜0.001）;移植时间（time）与移植方法（group）的交互效应显示不同的治疗方法时间趋势相同（F =35.54,P＞0.05）。大鼠神经功能评分组间比较显示,不同移植方法对大鼠神经功能评分变化的差异有显著性（F =6.434,P =0.001）;HUCBMC移植术后1天脑局部移植组大鼠神经功能评分（2.35±0.67）高于其他组,移植术后3、7、14天脑局部移植组大鼠神经功能评分（分别为0.40±0.60,0.25±0.37,0.03±0.22）低于其他组;HUCBMC移植后1天4组大鼠神经功能评分差异无显著性（F =2.14,P =0.10）;移植3、7、14天4组大鼠神经功能评分差异有显著性（F值分别为5.59,22.94,11.07,其对应P值均＜0.01）;相同移植途径不同时间点多个样本均数之间比较各组差异有显著性（F值分别为27.71,29.07,92.11,13.47,其对应P＜0.001）;尾静脉与左心室移植组各时间点神经功能评分差别无显著性（P分别为0.85,0.08,0.70,0.68）。结论 经尾静脉、左心室及脑出血局部途径移植人脐血单核细胞治疗脑出血大鼠神经功能均有改善;脑局部移植是脐血单核细胞移植的最佳途径。     

祛瘀开窍法治疗高血压脑出血术后脑水肿的临床疗效

目的探讨祛瘀开窍法治疗高血压脑出血术后脑水肿的临床疗效。方法选取我院2014-03-2014-09收治的高血压脑出血病人48例,随机分为实验组和对照组,实验组采取祛瘀开窍法联合西医综合治疗,对照组单用西医综合治疗,比较2组患者的术后脑水肿程度、格拉斯哥(GCS)评分、神经功能缺损评分及临床疗效。结果 2组术后24h、第3天脑水肿程度和GSC评分比较差异无统计学意义(P0.05),术后第7天、第14天组间比较,差异具有统计学意义(P0.05);术后24h、第14天神经功能缺损评分和疗效组间比较,实验组优于对照组,差异具有统计学意义(P0.05)。结论祛瘀开窍法治疗高血压脑出血术后的患者,可缓解脑水肿的程度,改善血流状态,提高患者临床疗效,值得推广。     

早期综合康复对脑出血术后瘫痪肢体功能恢复的作用

目的探讨早期综合康复对脑出血术后瘫痪肢体功能恢复的作用。方法对34例脑出血术后患者分组治疗护理,治疗组除常规药物治疗外,应用主、被动疗法,针灸、高压氧疗方法,并于3~4周进行肢体功能评定。结果治疗组肌力恢复有效率明显高于对照组,FMA和ADL评分明显高于对照组,2组比较差异有统计学意义(P<0.05)。结论早期综合康复能有效促进脑出血术后瘫痪肢体功能的恢复。     

辛伐他汀对局灶性脑缺血大鼠半暗带区缓激肽受体mRNA表达的影响

目的：探讨辛伐他汀预处理对局灶性脑缺血大鼠半暗带缓激肽受体基因表达的影响。方法：72只雄性SD大鼠随机分为3组：辛伐他汀干预组（干预组,给予辛伐他汀10 mg.kg-1.d-1和生理盐水1 mL的悬浊液灌胃）,脑缺血再灌注模型组（模型组,给予等体积生理盐水灌胃）,假手术组（给予等体积生理盐水灌胃）,3组分别预处理14 d。参照Longa法将干预组和模型组建立大脑中动脉栓塞模型,假手术组仅暴露右侧颈总和颈内动脉主干,不栓塞。并将各组随机分为再灌注3、24和48 h 3个亚组（均n=8）。于对应时间点行神经功能评分,用苏木精-伊红染色检测脑组织形态学,荧光定量RT-PCR技术检测脑缺血半暗带缓激肽B1、B2受体（BK-1Rs、BK-2Rs）的基因表达水平。结果：与模型组相比,3、24和48 h干预组大鼠缺血再灌注后神经功能改善、功能评分值降低（分别P〈0.05,P〈0.05,P〈0.01）,脑组织病理形态学变化减轻。荧光定量RT-PCR检测发现,3 h模型组脑缺血半暗带BK-1Rs、BK-2Rs与假手术组比表达减低,差异有统计学意义（P〈0.05,P〈0.01）;3 h干预组BK-1Rs、BK-2Rs与模型组比表达增加,差异有统计学意义（P〈0.01,P〈0.05）,24和48 h模型组脑缺血半暗带BK-1Rs与假手术组比表达减低,差异有统计学意义（P〈0.01,P〈0.01）;24和48 h干预组BK-1Rs与模型组比表达增加,差异有统计学意义（P〈0.01,P〈0.01）。结论：辛伐他汀预处理可改善大鼠局灶性脑缺血再灌注神经功能缺损及组织病理形态学,其机制可能与增加脑缺血半暗带BK-1Rs、BK-2Rs的表达有关。     

Module modified acute physiology and chronic health evaluation II: predicting the mortality of neuro-critical disease

Abstract

Objectives:

This study aimed to conduct and assess a module modified acute physiology and chronic health evaluation (MM-APACHE) II model, based on disease categories modified-acute physiology and chronic health evaluation (DCM-APACHE) II model, in predicting mortality more accurately in neuro-intensive care units (N-ICUs).

Methods:

In total, 1686 patients entered into this prospective study. Acute physiology and chronic health evaluation (APACHE) II scores of all patients on admission and worst 24-, 48-, 72-hour scores were obtained. Neurological diagnosis on admission was classified into five categories: cerebral infarction, intracranial hemorrhage, neurological infection, spinal neuromuscular (SNM) disease, and other neurological diseases. The APACHE II scores of cerebral infarction, intracranial hemorrhage, and neurological infection patients were used for building the MM-APACHE II model.

Results:

There were 1386 cases for cerebral infarction disease, intracranial hemorrhage disease, and neurological infection disease. The logistic linear regression showed that 72-hour APACHE II score (Wals = 173·04, P < 0·001) and disease classification (Wals = 12·51, P = 0·02) were of importance in forecasting hospital mortality. Module modified acute physiology and chronic health evaluation II model, built on the variables of the 72-hour APACHE II score and disease category, had good discrimination (area under the receiver operating characteristic curve (AU-ROC = 0·830)) and calibration (χ2 = 12·518, P = 0·20), and was better than the Knaus APACHE II model (AU-ROC = 0·778).

Discussion:

The APACHE II severity of disease classification system cannot provide accurate prognosis for all kinds of the diseases. A MM-APACHE II model can accurately predict hospital mortality for cerebral infarction, intracranial hemorrhage, and neurologic infection patients in N-ICU.     

大鼠脑出血致多器官功能障碍综合征模型的建立

目的:建立符合临床实际、简便易行的脑出血致多器官功能障碍综合征(MODS)的动物模型。方法:(1)采用两种剂量胶原酶+适量肝素注入大鼠尾状核构建脑出血模型,96只Wistar大鼠随机被分为正常对照组,假手术组,出血1组(胶原酶0.4u,肝素钠3.2IU),出血2组(胶原酶0.8u,肝素钠3.2IU),后2组又分成4h、8h、12h、24h、36h、48h和72h时相点的7个亚组,每组各6只大鼠:(2)记录大鼠脑出血后各时相点的症状、体征变化,检测外周血WBC、血浆内毒素、肝功、肾功、心肌酶学及各器官组织的病理变化,依据诊断标准判断全身炎症反应综合征(SIRS)、MODS的发生率。结果:(1)出血1组、2组动物的体温、呼吸、心率、WBC、血浆内毒素及各生化指标与正常组、假手术组相比有显著性差异(P<0.05)。出血2组与出血1组比较,上述指标亦存在明显统计学差异(P<0.05),尤其以外周血WBC、血浆内毒素差异更加明显(P<0.01)。(2)大鼠脑出血各时相点动物的器官组织均有不同程度的炎性损害,出血1组在24～36小时的脏器病理变化达到高峰,72小时基本恢复正常;出血2组较出血1组炎性损害更加明显持久,在24～48小时的脏器病理变化达到高峰,72小时仍可见炎性损害。(3)出血1组SIRS发生率75.4％,MODS发生率21.4％。出血2组SIRS发生率100％,MODS发生率67.9％。结论:以0.     

Heliox and oxygen reduce infarct volume in a rat model of focal ischemia

Normobaric hyperoxia treatment has recently been demonstrated to be remarkably beneficial in acute focal ischemia. The present study compared hyperoxia treatment with a novel heliox treatment. Adult male rats breathed 30% oxygen and 70% nitrogen (control group), 100% oxygen (hyperoxia group), or 30% oxygen and 70% helium (heliox group) during a middle cerebral artery occlusion for 2 h and a 1-hour reperfusion (n=6 in each group). Neurological deficits were scored at 3 and 24 h post focal ischemia. Neither the physiological parameters (body temperature, blood pressure, heart rate, O(2) saturation, and laser Doppler cerebral blood) nor the 3-hour post ischemia neurological scores differed between groups. However, the neurological scores showed a statistically significant improvement at 24 h post ischemia in the heliox group (p<0.05). The infarct volume (mean+SD) as measured by 2,3,5-triphenyltetrazolium staining included 36+/-17% of the involved hemisphere in the control group, 16+/-14% in the hyperoxia group, and 4+/-2% in the heliox group (p<0.01). In conclusion, whereas hyperoxia reduced the infarct volume, heliox further reduced the infarct volume and improved 24-hour neurological deficits in a rat model of focal ischemia. This suggests that a greater benefit may accrue from heliox therapy.     

地塞米松血肿腔内注入治疗大鼠脑出血的实验研究

目的研究地塞米松血肿腔内注入对大鼠脑出血灶周围水通道蛋白4(AQP4)、肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)含量变化的影响。方法应用自体尾动脉血注入法建立大鼠脑出血模型,54只SD大鼠,随机分为假手术组、脑出血模型组和地塞米松治疗组,每组18只,分别于48h、3d、7d断头取脑,每个时间点各6只,免疫组化染色观察脑出血灶周围AQP4、TNF-α、IL-10含量变化。结果①治疗组AQP4、TNF-α阳性细胞数百分比较脑出血组下降,差异具有统计学意义(P﹤0.05);②治疗组IL-10阳性细胞数百分比较脑出血组增高,差异具有统计学意义(P﹤0.05);③治疗组脑水肿含量较脑出血组下降,差异具有统计学意义(P﹤0.05)。结论地塞米松血肿腔内注射可以减少出血灶周围AQP4、TNF-α的表达,增加出血灶周围IL-10的表达。     

MMP-9抑制剂对脑出血大鼠血管再生的作用

目的 探讨基质金属蛋白酶-9（MMP-9）对脑出血大鼠血管再生的影响.方法 通过自体血注射制作大鼠脑出血模型,脑出血后第1～7天给予MMP-9抑制剂（盐酸多西环素30 mg/kg,1次/d,灌胃）进行干预,第7天运用Longa评分法进行神经功能评分后,处死动物并取脑,免疫组化法检测CD34抗体标记的血管内皮细胞,测定血肿周围微血管密度（MVD）,并测定脑出血后血肿周围侧脑室附近的室管膜下区（SVZ） MMP-9的表达的灰度值,对结果进行统计学分析.结果 与对照组相比,模型组MMP-9表达增加、MVD增加,而干预组较模型组MMP-9表达减少,MVD增加,差异有统计学意义（P＜0.05）.干预组较模型组神经功能评分降低,差异有统计学意义（P＜0.05）.结论 脑出血急性期大鼠抑制MMP-9表达可能对其恢复期血管再生及神经功能修复有促进作用.     

人工合成E-选择素对大鼠脑缺血再灌注损伤后血脑屏障的影响

目的探讨人工合成E-选择素对局灶性脑缺血再灌注损伤大鼠血脑屏障通透性以及紧密连接咬合蛋白和闭锁小带蛋白-1含量表达的影响。方法 180只SD大鼠随机分为假手术组(用线栓法将线栓插入颈内动脉及颈外动脉分叉处,其余操作同模型组)、模型组(用改良Zea Longa线栓法建立大鼠局灶性脑缺血再灌注损伤模型)、治疗组(建模前10 min从股静脉缓慢注射人工合成E-选择素10 mg·kg-1)。每组分为3、6、24、48和72 h 5个时间点(每个时间点大鼠n=12),应用多功能酶标仪测量血脑屏障对伊文思蓝(EB)的通透性,应用免疫组化法和Western blot法测定咬合蛋白及闭锁小带蛋白-1的表达含量。结果与假手术组比较,模型组缺血再灌注损伤后3 h脑组织EB含量开始上升,6 h继续上升,24 h达峰值,48~72 h下降但仍高于初始值;治疗组各时间点EB含量均低于模型组,两组比较差异有统计学意义(P0.05)。模型组咬合蛋白和闭锁小带蛋白-1的表达都于缺血再灌注3 h开始下降,6 h继续下降,24 h达最低值,48~72 h上升但低于初始值;治疗组各时间点咬合蛋白和闭锁小带蛋白-1的表达含量均高于模型组,两组比较差异有统计学意义(P0.05)。结论人工合成E-选择素可减轻大鼠脑缺血再灌注损伤造成的血脑屏障破坏,其机制可能与抑制紧密连接咬合蛋白和闭锁小带蛋白-1表达的减少有关。     

人工合成E-选择素对大鼠脑缺血再灌注损伤保护作用的实验研究

目的：探讨人工合成E-选择素对大鼠脑缺血再灌注损伤的保护作用机制。方法：雄性SD大鼠90只，随机分为3组：①假手术组；②缺血再灌注组；③人工合成B选择素治疗组（E-选择素治疗组）。大鼠局灶性脑缺血再灌注模型中B选择素治疗组建立模型前5min从股静脉注入人工合成E-选择素10mg·kg^-1。在不同时间点（缺血再灌注后2、6、12、24、48和72h）用ELISA法测定血浆IL-1β、TNF-α含量。分别在光镜和电镜下观察大鼠脑缺血再灌注区的病理形态改变。结果：缺血再灌注组与假手术组相比，血浆IL-1β、TNF-α含量明显增加（P〈0．05），E-选择素治疗组血浆IL-1β、TNF-α水平均降低（P〈0．05）。光镜和电镜下观察见缺血再灌注组额顶叶皮质和基底节区神经细胞呈缺血性改变，应用人工合成E-选择素后上述区域缺血性改变可明显减轻。结论：应用人工合成E-选择素能减轻大鼠脑缺血再灌注损伤后炎症细胞因子IL-1β和TNF-α的表达，减轻神经细胞缺血性改变，对大鼠脑缺血再灌注损伤具有保护作用。     

老龄大鼠急性脑缺血再灌注对心肌组织的影响

目的:研究老龄大鼠急性脑缺血再灌注对心肌组织的影响。方法:参照Zea Longa线栓法建立老龄大鼠局灶性脑缺血再灌注模型,从心肌酶谱、心电图及心肌超微结构等方面观察对心脏的影响。结果:与对照组比,在脑缺血2 h再灌注3和6 h组AST和LDH升高明显;CPK和CPK-MB在3、6和12 h组均有显著性升高(t检验,P<0.05)。大脑中动脉阻塞后出现ECG异常发生率为53.13%,6、12 h组明显高于32、4 h组(χ2检验,P<0.05),主要表现为S-T段的上抬和各种心律失常,在脑缺血30 min即可出现,70.59%发生在脑缺血后2 h和再灌注6 h内。心肌超微结构的改变在脑缺血2 h再灌注3和6 h组最明显,表现心肌缺血性损害和胞质中糖原颗粒明显减少。结论:急性脑缺血及再灌注后对心肌有明显损伤作用,以脑缺血2 h和再灌注6 h内最明显。     

补阳还五汤对脑出血大鼠PI3K/AKT信号通路的影响及其神经保护作用的机制

目的观察并探究补阳还五汤对脑出血大鼠脑组织磷脂酰肌醇3激酶/丝氨酸/苏氨酸蛋白激酶(PI3K/AKT)信号转导通路的影响及其神经保护作用的可能机制。方法 72只SD大鼠随机分为四组:假手术组、模型组、补阳还五汤组、银杏叶片组,每组18只。其中模型组、补阳还五汤组、银杏叶片组采用Rosenberg法制作脑出血大鼠模型。Garcia法检测大鼠神经功能评分,电镜观察神经元线粒体超微结构,western blot法检测磷酸化蛋白激酶(p-AKT)蛋白表达,TUNEL法检测细胞凋亡变化,免疫组化法检测B细胞淋巴瘤基因-2(bcl-2)蛋白、Bcl-2相关X蛋白(bax)的表达,甲酰胺法检测血脑屏障(BBB)通透性,干湿重法检测脑组织含水量。结果与脑出血模型组相比,补阳还五汤治疗明显提高神经功能学评分(P0.05),降低BBB通透性,减少脑组织水含量(P0.05),上调p-AKT、bcl-2蛋白表达,下调bax蛋白表达,减轻线粒体损伤,抑制神经元凋亡。结论补阳还五汤抗脑出血引起脑损伤的作用机制可能与其激活PI3K/AKT信号途径,抑制神经元凋亡,降低BBB通透性,减轻脑水肿有关。