VEGF与缺血缺氧性脑损伤

缺血缺氧可造成全身多个系统受损,尤其是中枢神经系统,缺血缺氧对脑损害最严重,血管内皮生长因子(VEGF)的血管生成作用、神经保护作用、神经再生作用能在一定程度上改善缺血缺氧性脑损伤,是一种潜在的缺血缺氧保护剂,本文就近年来VEGF在缺血缺氧性脑损伤方面的相关研究综述如下.     

亚低温减少沙土鼠脑缺血后延迟性神经元死亡机制的研究

目的：研究亚低温对脑缺血后延迟性神经元死亡的影响及其与海马羟自由基产生以及纹状体多巴胺和ＡＴＰ含量变化的关系。方法：沙土鼠前脑缺血再灌注模型,缺血１０ｍｉｎ,应用病理检查方法判断海马ＣＡｌ锥体细胞死亡的数目。动物随机分为假手术组、缺血组、缺血再灌注组和亚低温缺血再灌注组。高效液相加电化学检测器方法测定海马羟自由基和纹状体多巴胺的含量,高效液相紫外检测器法测定纹状体ＡＴＰ含量。结果：亚低温条件下沙土     

IL-1β在缺血致脑损伤中的主要作用机制

中枢白细胞介素-1β（IL-1β）在缺血引起的脑损伤过程中发挥十分重要的作用,但其作用机制尚不十分明确,本文主要就近年来的研究进展,从其对诱导粘附分子表达,激活内皮细胞产生多种活性物质,促进脑内兴奋性氨基酸和自由基等神经毒性物质的产生和释放等角度,简要概括其在缺血引起的脑损伤过程中的作用机制。     

神经节苷脂与头部贴敷式亚低温联合治疗对新生鼠缺氧缺血性脑损伤的影响

目的:研究神经节苷脂与头部贴敷式亚低温联合治疗新生鼠缺氧缺血性脑损伤时脑组织内一氧化氮(NO)和(MDA)含量的变化,为新生儿缺氧缺血性脑病的治疗提供理论依据.方法:将88只WISTAR新生鼠随机分为四组,神经节苷脂治疗组(IG)、头部贴敷式亚低温治疗组(IH)、联合治疗组(IB)以及对照组(CN).结果:CN组NO和MDA的含量明显升高,IG、IH以及IB组NO和MDA均明显下降.结论:神经节苷脂和亚低温通过降低NO和MDA的含量保护缺氧缺血神经元,二者联合效果更好.     

梓醇对大鼠脑缺血再灌注损伤的保护作用研究

目的:探讨梓醇对缺血再灌注大鼠脑损伤后的保护作用.方法:采用传统大脑中动脉阻塞(MCAO)方法制备大鼠局灶性缺血模型,根据随机数字表法将SD大鼠分为MCAO组、对照组(vehicle组)及梓醇处理组(catalpol组),缺血再灌注48 h后观察各组大鼠神经功能学评分和脑梗死容积.分别于术前、术后6h、24 h、48 h取大鼠脑组织样本,检测匀浆中谷胱甘肽过氧化物酶(GSH-PX)和丙二醛(MDA)的变化情况.结果:与vehicle组和MCAO组相比,catalpol处理组神经功能学评分降低(P＜0.05);其梗死容积较小(P＜0.05).组织匀浆结果显示catalpol处理组脑匀浆中GSH-PX活力升高,MDA含量下降(P＜0.05).结论:梓醇可能通过降低脑内自由基水平、控制脂质过氧化程度,对缺血再灌注引起的大鼠脑损伤产生神经保护作用.     

人参与丹参的抗脑缺氧作用

中药人参与丹参在我国的药用历史都很久。一些研究表明 ,人参制剂有抗缺氧作用 ,人参的主要活性成份人参皂甙具有抗缺氧性脑损伤的作用。同样 ,丹参亦具有抗缺氧及抗缺氧性脑损伤的作用 ,并且临床上已用于缺血性心脑血管疾病等治疗。然而 ,人参与丹参抗缺氧脑损伤的作用机制目前尚不十分清楚 ,人参亦尚未用于临床防治缺氧缺血性脑病。本课题结合整体动物及离体海马脑片的低氧实验 ,以比较人参与丹参的抗脑缺氧作用 ,并分析其作用机制 ,为临床应用人参防治缺血缺氧性脑损伤提供理论依据。1　材料和方法(1)药剂　人参及丹参制剂由上海海洋水下…     

异丙酚对全脑缺血/再灌注大鼠海马谷氨酸、抗坏血酸释放的影响

目的：观察异丙酚对全脑缺血／再灌注大鼠海马细胞外谷氨酸（Glu）和抗坏血酸（AA）的影响,探讨异丙酚脑保护作用机制。方法：采用Pulsinelli-Brlerley四血管阻断法制备全脑缺血模型,应用脑微透析技术结合高效液相色谱（HPLc）检测大鼠海马细胞外Glu、AA含量的变化。结果：与缺血／再灌注组各对应时点相比较,异丙酚处理组大鼠海马细胞外Glu、AA含量明显降低,统计结果差异均有显著性（P〈0．05,或〈0．01）。结论：缺血／再灌注早期应用异丙酚不仅减少兴奋性氨基酸释放,还能清除自由基、抑制脂质过氧化反应而产生脑保护作用。     

头颅局部亚低温对脑梗死治疗作用的临床研究

目的：观察头颅局部行亚低温治疗脑梗死后患者血清一氧化氮、内皮素以及细胞间粘附分子含量的变化,探讨亚低温治疗对脑组织发挥保护作用的机制。方法：搜集2012年5月至2013年2间,哈尔滨医科大学附属第四医院神经内科收治的62例脑梗死患者,并随机分为亚低温治疗组（32例）和常规治疗对照组（30例）,治疗组在药物治疗脑梗死的同时加用局部亚低温治疗技术,在治疗前、治疗后3、7、14d进行一氧化氮、内皮素及细胞间粘附分子的测定;两组患者分别于治疗前以及治疗后第30d,对神经功能缺损采用NIHSS量表进行评分,对日常生活活动能力采用Bathel指数评分,对运动功能采用FuglMeyer法进行评价。结果：在治疗后第3、7、14天,亚低温治疗组一氧化氮含量较对照组显著升高（均为P〈0．01）,在治疗后第7天亚低温治疗组内皮素含量低于对照组（P〈0．05）,第3、14天亚低温组内皮素含量显著低于对照组（P〈0．01）;治疗前两组NIHSS评分、Bamel指数和FuglMeyer评分差异均无显著性意义（P〉0．05）。在治疗组治疗后的第3、7、14天,亚低温治疗组细胞间粘附分子与治疗前相比,均出现降低（P〈0．05）;而且在第3天,治疗组和对照组有显著性差异（P〈0．05）。治疗1个月后,亚低温治疗组NIHss评分比常规治疗组显著降低,亚低温治疗组Bathel指数和FuglMeyer评分均明显高于对照组,差异具有显著性。结论：局部亚低温治疗对血管内皮细胞功能有较好的保护作用,可改善脑梗死患者神经功能缺损及预后。     

新生儿缺血缺氧性脑病发病机理及其治疗方法

新生儿缺血缺氧性脑病 (HIE)主要是指新生儿在围产期窒息导致的缺血缺氧性损害 ,临床上出现一系列脑病的表现。 HIE至今仍是威胁新生儿生命和健康的严重疾病。近年来 ,国内外对此病的研究较多 ,现就其发病机理及其治疗方法综述如下。1　发病机理　　新生儿缺血缺氧性脑病的发病原因很多 ,目前认为主要是围产期窒息 ,其发病机理大多认为与氧自由基、Ca2 内流及再灌注有关。1 .1　 HIE与氧自由基　正常生物体内有少量自由基产生 ,并经清除系统不断清除 ,从而维持动态平衡 ,使机体免遭损害。当机体自由基浓度高于正常水平时 ,自由基清除系…     

孕酮对新生大鼠低氧缺血性脑损伤中MMP-3表达的影响

目的：研究孕酮（PROG）对新生大鼠低氧缺血后脑内基质金属蛋白酶3（MMP-3）表达的影响。方法：建立新生大鼠低氧缺血性脑损伤动物模型,伊文思兰（EB）染色和电镜观察新生鼠低氧缺血性脑损伤血一脑屏障的通透性改变;免疫印迹（Western blot）方法检测大脑皮层MMP-3表达。结果：电镜显示低氧缺血组血-脑屏障完整性明显破坏：EB染色结果表明低氧缺血组血-脑屏障通透性明显高于假手术组,差异极显著（P〈0．01）,孕酮组血-脑屏障通透性明显低于低氧缺血组,有显著性差异（P〈0．05）;Western blot结果显示低氧缺血组MMP-3蛋白表达显著高于假手术组（P〈0．01）;孕酮组MMP-3蛋白表达显著低于低氧缺血组（P〈0．05）。结论：孕酮通过减少MMP-3的表达,降低血一脑屏障的损伤,这可能是其发挥脑保护作用的机制之一。     

Dynamics of free amino acids in rat brain tissue during hypothermia

A comparative study is conducted for the effect of one-, three- and six-hour artificial deep (20-19 degrees C) hypothermia on the content of free amino acids in the blood serum, tissue, nuclei and mitochondria of the rat brain. It is found out that the content is the highest in the blood serum after a three-hour cooling. In the brain tissue the amount of amino acids lowers, especially under conditions of a six-hour hypothermia. In nuclei a three-hour effect of hypothermia decreases sharply the content of free amino acids and the six-hour one increases the amount of most of them. Under hypothermia the content of nearly all amino acids in the brain mitochondria is higher than in the intact animals.     

Effect of mild hypothermia on blood brain barrier disruption induced by oleic acid in rats

The blood-brain barrier (BBB) is essential for the normal function of the central nervous system. The pathological conditions induced by brain diseases including cerebral ischemia result in the alteration of BBB integrity. This alteration of BBB is relieved by mild hypothermia that has been regarded as an effective therapy for brain injury. Experimental fat embolism by intra-arterial administration of fatty acid induces reversible dysfunction of BBB and is considered as a beneficial method for the research on BBB disruption. However, the implication of hypothermia on the fatty acid-induced BBB disruption is not clear yet. In this study, we aim to investigate the effect of mild hypothermia on BBB disruption by comparing the changes of brain inflammation, free radical production, and matrix metalloproteinases (MMPs) caused by cerebral fatty acid infusion between normothermic (37°C) and hypothermic (33°C) groups. Oleic acid infusion into the carotid artery induced the increase of BBB permeability, which was inhibited by mild hypothermia. Neutrophils were infiltrated and intercellular adhesion molecule-1 (ICAM-1) expression was increased in the vascular structures in the affected brain tissue of normothermic rats at 24 hrs following oleic acid administration. Inducible nitric oxide synthase (iNOS) and nitro-tyrosine immunoreactivities were also observed in the normothermic group. The expression of matrix metalloproteinase (MMP)-2, 3, and 13 were upregulated predominantly in the oleic acid-treated brain of the normothermic rats. In mild hypothermic condition, neutrophil infiltration and ICAM-1 expression were attenuated, whereas the inductions of iNOS, nitrotyrosine and MMPs except MMP3 were not affected. Therefore, we suggest that mild hypothermia contributes to the protective effect on oleic acid-induced BBB damage via reducing neutrophil infiltration and brain inflammation.     

Effects of postdecapitative ischemia and hypoxia on the phosphoglyceride acyl groups of rat brain membranes

Rats subjected to mild hypoxic and postdecapitative ischemic treatments indicated a decrease (8–16%) in the proportion of polyunsaturated acyl groups of diacyl glycerophosphocholines (diacyl-GPC), diacyl glycerophosphoethanolamines (diacyl-GPE), and alkenylacyl glycerophosphoethanolamines (alkenylacyl-GPE) in brain synaptosomes. In general, the acyl group changes due to mild hypoxic treatment were less obvious than those due to the ischemic treatment. The decrease in polyunsaturated acyl groups was marked by an increase in the saturated (16:0 and 18:0) and monoenoic (18:1) acyl groups. Among the polyunsaturated acyl groups, arachidonate (20:4) indicated the greatest decrease in response to ischemic and hypoxic treatments. The decrease in polyunsaturated fatty acids of diacyl glycerophosphocholines was largest in the first minute of ischemic treatment and the first 30 min of hypoxic treatment. After the initial decrease, there was a slight recovery. The biphasic type of change was thought to be due to active reacylation of the lyso phospholipids. This biphasic change, however, was not observed with ethanolamine phosphoglycerides which indicated a steady decrease in the polyunsaturated acyl group content with time of ischemic treatment. The increased hydrolysis of polyunsaturated acyl groups in brain membrane phosphoglycerides due to the ischemic and hypoxic treatments seemed to correlate well with the implication of phospholipase A₂ involvement in eliciting the increase in free fatty acids during brain stimulation.     

Effect of Cooling on Force-Frequency Relationship,Rest Potentiation,and Frequency-Dependent Acceleration of Relaxation in the Guinea Pig Myocardium

Journal of Evolutionary Biochemistry and Physiology - While deep hypothermia is well known to lead to cardiac malfunction up to circulatory arrest, mild hypothermia can prevent hypoxic damage to...     

The Effect of Hypothermia Therapy on Cortical Laminar Disruption following Ischemic Injury in Neonatal Mice

Hypothermia has been proposed as a treatment for reducing neuronal damage in the brain induced by hypoxic ischemia. In the developing brain, hypoxic ischemia-induced injury may give rise to cerebral palsy (CP). However, it is unknown whether hypothermia might affect the development of CP. The purpose of this study was to investigate whether hypothermia would have a protective effect on the brains of immature, 3-day old (P3) mice after a challenge of cerebral ischemia. Cerebral ischemia was induced in P3 mice with a right common carotid artery ligation followed by hypoxia (6% O₂, 37°C) for 30 min. Immediately after hypoxic ischemia, mice were exposed to hypothermia (32°C) or normothermia (37°C) for 24 h. At 4 weeks of age, mouse motor development was tested in a behavioral test. Mice were sacrificed at P4, P7, and 5 weeks to examine brain morphology. The laminar structure of the cortex was examined with immunohistochemistry (Cux1/Ctip2); the number of neurons was counted; and the expression of myelin basic protein (MBP) was determined. The hypothermia treatment was associated with improved neurological outcomes in the behavioral test. In the normothermia group, histological analyses indicated reduced numbers of neurons, reduced cortical laminar thickness in the deep, ischemic cortical layers, and significant reduction in MBP expression in the ischemic cortex compared to the contralateral cortex. In the hypothermia group, no reductions were noted in deep cortical layer thickness and in MBP expression in the ischemic cortex compared to the contralateral cortex. At 24 h after the hypothermia treatment prevented the neuronal cell death that had predominantly occurred in the ischemic cortical deep layers with normothermia treatment. Our findings may provide a preclinical basis for testing hypothermal therapies in patients with CP induced by hypoxic ischemia in the preterm period.     

亚低温联合镁剂对脑缺血再灌注大鼠保护作用的研究

目的:rt-PA溶栓为缺血性卒中最有效的治疗方法,脑血流再通后挽救濒临死亡的神经细胞同时,也可能发生更为严重而持久的脑缺血再灌注损伤。本研究探讨联合应用局部亚低温(32-35℃)及硫酸镁对局灶性脑缺血再灌注大鼠的保护作用及其可能机制。方法:通过线栓法建立大鼠大脑中动脉阻塞(MCAO)及再通模型,将50只雄性Wistar大鼠随机分为假手术组、常温组、亚低温组、硫酸镁组、亚低温+硫酸镁组,每组10例,采用Longa神经功能评分、TTC染色、干湿重法、TUNEL技术,检测和比较各组脑缺血再灌注后大鼠的神经功能、脑梗死体积、脑组织含水量及凋亡细胞数。结果:与常温组相比,亚低温组与亚低温+硫酸镁组的梗死体积、神经功能评分、脑组织含水量、凋亡细胞数均明显降低,差异有显著意义(P0.05);而与亚低温组相比,亚低温+硫酸镁组局灶脑缺血大鼠的脑梗死体积、神经功能评分、脑组织含水量、凋亡细胞数均显著减少,差异有显著意义(P0.05)。结论:与单独应用亚低温相比,局部亚低温与硫酸镁联合应用,对局灶性脑缺血再灌注大鼠可发挥更有效的脑保护作用。其机制可能与抑制脑缺血再灌注后凋亡及减轻脑水肿有关。二者联用可能为缺血性卒中患者提供一种减轻溶栓后再灌注损伤的有效脑保护方法。     

褪黑素抑制低氧引起大鼠大脑皮层氨基酸递质释放

为研究褪黑素对低氧引起大鼠大脑皮层脑片氨基酸释放变化的影响,利用反相高效液相色谱结合荧光检测法,测定了孵育液中氨基酸类神经递质的含量。低氧条件为通入９１．６％Ｎ２和８．４％Ｏ２的混合气体。低氧３０ｍｉｎ时,大鼠大脑皮层脑片孵育中,氨基酸类神经递质天冬氨酸、谷氨酸、谷氨酰胺、甘氨酸、牛磺酸和γ－氨基丁酸的含量显著增加,其含量分别是正常氧组的２４０．４％,３３４．３％,２００．６％,２１０．４％,１６     

Reduction of ultraviolet light-induced oxidative stress by amino acid-based iron chelators

The generation of free radicals by ultraviolet (UV) light accelerates skin aging, which is known as photoaging. Cutaneous iron catalyzes the generation of free radicals. We designed novel antioxidants that suppressed the iron-catalyzed free radical generation and the ensuing UV-induced damage by mimicking the binding site of iron sequestering proteins. These antioxidants, N-(2-hydroxybenzyl)amino acids, were prepared by condensation of amino acids such as glycine and L-serine with salicylaldehyde and followed by catalytic reduction. The compounds formed a 2:1 complex to iron ion. These amino acid derivatives inhibited the iron-induced hydroxyl radical generation (the Fenton reaction). The compounds also suppressed UV-induced lipid peroxidation in murine dermal fibroblast homogenates. In addition, N-(2-hydroxybenzyl)-L-serine showed protective activity against UV-induced cytotoxicity in murine dermal fibroblasts. Desferrioxamine, a strong iron sequestering compound, was effective in inhibiting the Fenton reaction and the lipid peroxidation, but it was ineffective in protecting against UV-induced cytotoxicity. The results suggest that UV-induced oxidative stress can be reduced by these amino acid derivatives.     

The effect of a free radical scavenger and platelet-activating factor antagonist on FFA accumulation in post-ischemic canine brain

The effects of the platelet-activating factor antagonist BN 50739 and a free radical scavenger dimethyl sulfoxide on the accumulation of free fatty acids in post-ischemic canine brain are reported. Following 14 min of complete normothermic ischemia and 60 min of reperfusion, the total brain FFAs were approximately 150% higher than in the control group (p<0.05). Perfusion with the platelet-activating factor antagonist BN50739 in its diluent dimethyl sulfoxide during 60 min of post-ischemic reoxygenation resulted in a 61.8% (p<0.01) reduction in the total brain free fatty acid accumulation. Palmitic, stearic, oleic, linoleic, and arachidonic acids decreased by 53.8%, 63.5%, 69.0%, 47.4%, and 57.2%, respectively. Although dimethyl sulfoxide alone caused stearic and arachidonic acids to return to the normal concentration range, BN 50739 had a significant influence on recovery of palmitic, oleic, and linoleic acids and was previously shown to provide significant therapeutic protection against damage to brain mitochondria following an ischemic episode. Because free fatty acid accumulation is one of the early phenomena in cerebral ischemia, this study provides evidence to support the hypothesis that both platelet-activating factor and free radicals are involved in initiating cerebral ischemic injury.     

Separation, detection, and quantification of hydroperoxides formed at side-chain and backbone sites on amino acids, peptides, and proteins

Hydroperoxides are major reaction products of radicals and singlet oxygen with amino acids, peptides, and proteins. However, there are few data on the distribution of hydroperoxides in biological samples and their sites of formation on peptides and proteins. In this study we show that normal-or reversed-phase gradient HPLC can be employed to separate hydroperoxides present in complex systems, with detection by postcolumn oxidation of ferrous xylenol orange to the ferric species and optical detection at 560 nm. The limit of detection (10-25 pmol) is comparable to chemiluminescence detection. This method has been used to separate and detect hydroperoxides, generated by hydroxyl radicals and singlet oxygen, on amino acids, peptides, proteins, plasma, and intact and lysed cells. In conjunction with EPR spin trapping and LC/MS/MS, we have obtained data on the sites of hydroperoxide formation. A unique fingerprint of hydroperoxides formed at alpha-carbon (backbone) positions has been identified; such backbone hydroperoxides are formed in significant yields only when the amino acid is part of a peptide or protein. Only side-chain hydroperoxides are detected with free amino acids. These data indicate that free amino acids are poor models of protein damage induced by radicals or other oxidants.