Chinese herbal extract dl-3n-butylphthalide A commonly used drug for the treatment of ischemic stroke as a novel therapeutic approach to treat neurodegenerative diseases

Dl-3n-butylphthalide is the active component isolated from the seeds of Apium graveolens Linn.A number of pharmacological and clinical studies have proven that dl-3n-butylphthalide is highly potent and multi-targeted with low toxicity and has a long time-window for the treatment of ischemic cerebrovascular disease.The mechanisms underlying dl-3n-butylphthalide include improving mi-tochondrial function and microcirculation,inhibiting apoptosis and reducing oxidative stress.Fur-thermore,dl-3n-butylphthalide may also be promising for the treatment of neurodegenerative dis-eases,such as Alzheimer’s disease,vascular dementia and Parkinson’s disease.     

VEGF-165基因对创伤性脑损伤脑血流动力学影响的实验研究

目的 研究外源性血管内皮细胞生长因子(VEGF)基因治疗大鼠创伤性脑损伤(TBI)后脑灌注的变化,了解其血流动力学改变.方法 创伤性脑损伤大鼠模型建立后随机分为3组:治疗组,质粒对照组,外伤组.通过RT-PCR检测脑伤后1h、6h、24h、3d、7d、14 d VEGF mRNA在损伤局部的表达改变;应用CT灌注像(CTP)研究不同时间脑血流量(CBF)、脑血容量(CBV)等参数在VEGF-165基因治疗前后的动态变化.结果 VEGF-165基因治疗创伤性脑损伤大鼠后经RT-PCR扩增的VEGF mRNA绝对积分光密度值水平显著高于外伤组和质粒对照组(P＜0.05).CTP参数和伪彩图均显示基因治疗组脑灌注在伤后24h CBF、CBV有增高趋势,伤后3d、7d脑灌注明显高于TBI 组(P＜0.05),虽然伤后14 d CBF、CBV开始降低,但和TBI组比较仍然较高.结论 本研究结果显示大鼠创伤性脑损伤后外源性VEGF基因能够提高脑损伤组织的脑灌注,改善脑损伤部位微循环,为损伤组织的恢复提供基础.     

丁基苯酞对脑出血大鼠血管内皮生长因子、血管生成素-2蛋白表达及血管新生的影响

目的 通过观察大鼠脑出血后脑组织中血管内皮生长因子（vaseular endothelial growth factor, VEGF）、血管生成素-2（angiopoietin-2,Ang-2）蛋白在不同时间点的动态表达及不同剂量丁基苯酞干 预后其与新生血管数量的变化,探讨丁基苯酞对脑出血大鼠可能的神经保护作用及机制。 方法 通过自体血注入法制备SD大鼠脑出血模型并随机分为脑出血模型组、丁基苯酞低剂量组 及丁基苯酞中剂量组,以及对照的假手术组。丁基苯酞低、中剂量组大鼠分别予以丁基苯酞10 mg/kg、 25 mg/kg灌胃给药（每日2次）,假手术组及脑出血模型组大鼠则在相同时间用同等体积大豆油代替 灌胃。分别在术后1 d、3 d、7 d、15 d评估大鼠神经功能缺损,采用免疫组化法检测各时间点CD34的表 达并进行新生血管计数及血管场面积测定,检测各时间点VEGF、Ang-2蛋白的表达,测定脑出血大鼠 血肿体积。 结果 与脑出血模型组比较,丁基苯酞中剂量组在术后各时间点,低剂量组术后7 d、15 d VEGF蛋 白表达上调;丁基苯酞低、中剂量组术后各时间点An g-2蛋白表达均上调,血管计数均增多;丁基苯酞 低剂量组术后3 d的神经功能缺损评分较低,丁基苯酞中剂量组术后3 d、7 d、15 d的神经功能缺损评 分较低,上述差异均有统计学意义。丁基苯酞低、中剂量组与脑出血模型组,在术后7 d、15 d血肿体 积差异无统计学意义。 结论 丁基苯酞可以显著减轻脑出血大鼠的神经功能缺损,其作用机制可能与上调VEGF、Ang-2蛋 白的表达,增加脑出血血肿周围新生血管密度有关,同时未增加血肿增大的风险。     

丁苯酞对脑缺血再灌注损伤大鼠MMP-9活性和血脑屏障通透性的影响

目的探讨丁苯酞对脑缺血再灌注损伤大鼠的保护作用。方法通过线栓法制备大鼠局灶性脑缺血再灌注损伤模型,腹腔注射丁苯酞,于脑缺血再灌注后6h、12h、24h后用免疫组织化学的方法观察损伤侧脑组织MMP-9活性及基底膜成分Ⅳ型(collagenⅣ)胶原表达的变化。同时测定伊文思兰的含量观察损伤侧脑组织血脑屏障通透性。结果随缺血再灌注时间延长,MMP-9活性、EB含量逐渐增加,24h达峰,Ⅳ型胶原的表达逐渐减少,各组之间表达均具有统计学意义(P<0.05);丁苯酞处理组MMP-9活性、EB含量各个时间点明显低于缺血再灌注组,相同时间点Ⅳ型胶原的表达增加(P<0.05)。结论丁苯酞可抑制脑缺血再灌注损伤大鼠MMP-9的表达,增加基底膜Ⅳ型胶原的表达,降低血脑屏障通透性,从而发挥脑保护作用。     

大鼠流体冲击致脑损伤后海马循环的变化

目的 :探讨大鼠闭合性脑损伤后 ,脑循环功能的改变规律。方法 :应用流体冲击致大鼠脑损伤模型 ,采用氢清除法和二胺基联苯胺染色及图像分析 ,观察大鼠海马血流量及海马血管密度的变化。结果 :大鼠脑损伤后 1h海马血流量开始下降 ,4h达最低 ,8h后逐渐恢复 ;与此同时海马血管密度在伤后 4h升高达峰值 ,8h后渐下降至伤前水平。结论 :大鼠流体冲击致脑损伤后 ,海马血流量的降低与血管密度破坏性的增加表明脑循环明显受损     

白花丹参对促脑缺血再灌注后神经元再生的影响

摘要：目的：通过白花丹参干预大鼠局灶性脑缺血再灌注动物模型,探讨白花丹参对脑缺血再灌注后脑细胞再生及再生神经细胞分化的影响及影响机制。方法：采用线栓法制备大鼠局灶性脑缺血再灌注模型,给予白花丹参干预,分别从脑血流量、脑细胞凋亡率、新生脑细胞数量及新生神经细胞的分化等指标,观察白花丹参的脑保护作用及保护机制。结果：白花丹参具有增加脑血流量、降低脑细胞凋亡率、促进脑细胞再生及再生神经细胞的分化的作用。结论：缺血性脑损伤时白花丹参能显著的再加脑血流量、降低脑细胞凋亡率、促进脑细胞再生及再生神经细胞的分化。     

盐酸纳络酮对兔酒精中毒后脑外伤早期脑血流的影响

目的研究盐酸纳络酮(NAL)对兔急性酒精中毒后脑外伤早期脑血流动力学及脑血管形态变化的影响。方法家兔20只,随机分为NAL治疗组和生理盐水对照组,每组10只。乙醇灌胃法致使家兔急性酒精中毒,自由落体直接打击颅骨法制作实验性颅脑损伤模型。治疗组在伤后30min静注NAL2mg/kg,以后每隔60min重复注射一次。采用经颅多普勒(TCD)超声结合脑血管造影等方法,于伤前及伤后一定时间测定平均动脉压(MABP)、颅内压(ICP)、大脉中动脉舒张期流速(Vd)、脉搏指数(PI)、脑血管直径指数(CVI),进行统计学分析。结果两组动物伤后MABP明显下降,ICP及PI值显著升高,Vd明显减慢,CVI显著减小。NAL治疗后与治疗前及对照组各相对时间比较,MABP显著升高(P<0.05),Vd显著增快(P<0.01),PI值明显降低(P<0.01),CVI明显增大(P<0.05),TCD频谱接近正常。结论NAL(2mg/kg)可逆转酒精中毒所致的兔颅脑损伤后发生的显著的低血压及脑血管收缩、并可降低脑血管阻力,改善微循环,增加脑血流量,对兔急性酒精中毒后脑外伤有治疗作用。     

Sequelae following traumatic brain injury: The cerebrovascular perspective

Traumatic brain injury (TBI) initiates a huge repertoire of biochemical perturbations. On one hand, destructive events are set into motion while on the other hand, protective and recovery mechanisms are evoked, each with their own temporal and spatial characteristics. The brain exists as a finely tuned balance between vascular, neuronal and glial interactions and so a complex interplay between these factors will dictate the final evolution of pathogenesis. Although vascular damage is a key event, it remains a somewhat neglected component to the underlying degenerative processes that evolve following injury to the brain. The present review will act to integrate the current knowledge of the vascular events proceeding injury to the brain, with an emphasis on how this impacts the control of vascular function and thus cerebral blood flow.     

丁基苯酞加强造血干细胞移植及内源性干细胞动员治疗脑梗死的效应

Exogenous stem cell transplantation and endogenous stem cell mobilization are both effective for the treatment of acute cerebral infarction. The compound dl-3-butylphthalide is known to improve microcirculation and help brain cells at the infarct loci. This experiment aimed to investigate the effects of dl-3-butylphthalide intervention based on the transplantation of hematopoietic stem cells and mobilization of endogenous stem cells in a rat model of cerebral infarction, following middle cerebral artery occlusion. Results showed that neurological function was greatly improved and infarct volume was reduced in rats with cerebral infarction. Data also showed that dl-3-butylphthalide can promote hematopoietic stem cells to transform into vascular endothelial cells and neuronal-like cells, and also enhance the therapeutic effect on cerebral infarction by hematopoietic stem cell transplantation and endogenous stem cell mobilization.     

The role of opioids in newborn pig fluid percussion brain injury

The present study was designed to characterize the relationship between cerebral opioid concentration, cerebral hemodynamics, and cerebral oxygenation following percussion brain injury in neonatal pigs. Previous research found that opioids represent a significant vasoactive component in the regulation of the neonatal piglet cerebral circulation. Anesthesized newborn (1–5 days old) pigs equipped with a closed cranial window were connected to a percussion device consisting of a saline-filled cylindrical reservoir with a metal pendulum. Brain injury of moderate severity (1.9–2.3 atm.) was produced by allowing the pendulum to strike a piston on the cylinder. Fluid percussion brain injury decreased pial arteriolar diameter (132 ± 5 to 110 ± 5 μm within 10 min). Cerebral blood flow also fell within 10 min of injury and continued to fall progressively for 3 h, resulting in a 46 ± 4% decrease. Within 30 s of brain injury, there was a transient increase in cerebral hemoglobin-O₂ saturation that was reversed to a progressive profound decrease in cerebral hemoglobin-O₂ saturation for the next 3 h, as measured by near infrared spectroscopy. CSF opioid concentrations were increased 10 min after brain injury; dynorphin showed the largest proportional increase (5.8 ± 0.9 fold). The CSF concentration for other opioids continued to increase over 180 min while the dynorphin concentration progressively decreased with time. In naloxone (1 mg/kg i.v.) pretreated piglets, the brain injury induced decrease in arteriolar diameter was attenuated (129 ± 5 to 121 ± 5 μm within 10 min). Similarly, the decrease in regional cerebral blood flow and cerebral hemoglobin-O₂ saturation observed following brain injury were also blunted by naloxone. These data show that CSF opioid concentrations increase following brain injury and that the time course and relative increase in CSF concentration vary from opioid to opioid. These data also indicate that in the immature animal, opioids contribute to arteriolar constriction, decreased cerebral blood flow, decreased cerebral oxygenation, and could play a role in causing ischemia after brain injury.     

经颅多普勒超声在重型颅脑创伤救治中的临床应用

重型颅脑创伤（sTBI）的临床监测手段多以体格检查、影像学检查为主,而这些手段无法迅速有效地诊断、监测sTBI病理生理过程。经颅多普勒超声（TCD）可以无创、实时地测量大脑大动脉的血流状态,获得脑血流动力学信息,通过分析脑血流速度及方向、血管自身调节功能、远端血管阻力变化来监测脑血管狭窄、痉挛程度并评估颅内压、识别脑死亡。本文围绕TCD在sTBI救治中的临床应用综述如下。     

How does the motor relearning program improve neurological function of brain ischemia monkeys?

The motor relearning program can significantly improve various functional disturbance induced by ischemic cerebrovascular diseases. However, its mechanism of action remains poorly understood. In injured brain tissues, glial fibrillary acidic protein and neurofilament protein changes can reflect the condition of injured neurons and astrocytes, while vascular endothelial growth factor and basic fibroblast growth factor changes can indicate angiogenesis. In the present study, we induced ischemic brain injury in the rhesus macaque by electrocoagulation of the M1 segment of the right middle cerebral artery. The motor relearning program was conducted for 60 days from the third day after model establishment. Immunohistochemistry and single-photon emission CT showed that the numbers of glial fibrillary acidic protein-, neurofilament protein-, vascular endothelial growth factor- and basic fibroblast growth factor-positive cells were significantly increased in the infarcted side compared with the contralateral hemisphere following the motor relearning program. Moreover, cerebral blood flow in the infarcted side was significantly improved. The clinical rating scale for stroke was used to assess neurological function changes in the rhesus macaque following the motor relearning program. Results showed that motor function was improved, and problems with consciousness, self-care ability and balance function were significantly ameliorated. These findings indicate that the motor relearning program significantly promoted neuronal regeneration, repair and angiogenesis in the surroundings of the infarcted hemisphere, and improve neurological function in the rhesus macaque following brain ischemia.     

Galectin-3 protects against ischemic stroke by promoting neuro-angiogenesis via apoptosis inhibition and Akt/Caspase regulation

Post-stroke neurological deficits and mortality are often associated with vascular disruption and neuronal apoptosis. Galectin-3 (Gal3) is a potent pro-survival and angiogenic factor. However, little is known about its protective role in the cerebral ischemia/reperfusion (I/R) injury. We have previously shown significant up-regulation of Gal3 in the post-stroke rat brain, and that blocking of Gal3 with neutralizing antibody decreases the cerebral blood vessel density. Our current study demonstrates that intracerebral local delivery of the Gal3 into rat brain at the time of reperfusion exerts neuroprotection. Ischemic lesion volume and neuronal cell death were significantly reduced as compared with the vehicle-treated MCAO rat brains. Gal3 increased vessel density and neuronal survival after I/R in rat brains. Importantly, Gal3-treated groups showed significant improvement in motor and sensory functional recovery. Gal3 increased neuronal cell viability under in vitro oxygen–glucose deprivation conditions in association with increased phosphorylated-Akt, decreased phosphorylated-ERK1/2, and reduced caspase-3 activity. Gene expression analysis showed down regulation of pro-apoptotic and inflammatory genes including Fas-ligand, and upregulation of pro-survival and pro-angiogenic genes including Bcl-2, PECAM, and occludin. These results indicate a key role for Gal3 in neuro-vascular protection and functional recovery following ischemic stroke through modulation of angiogenic and apoptotic pathways.     

弥漫性脑损伤微血管形态学研究

目的 运用形态计量学方法研究弥漫性脑损伤（DBI）大鼠模型的微循环及胶质细胞形态学改变，并观察萘呋胺酯（NF）对DBI的治疗作用。方法 采用Marmarou的DBI模型，光镜下测量顶叶皮质内微血管及其周围水肿区的截面面积，电镜下半定量测定皮质区毛细血管周的水肿范围及胶质细胞胞体的肿胀程度。结果 外伤后2、6、24小时平均微血管截面面积与对照组比较无显著缩小，6、24小时微血管周水肿明显增加。NF组伤后2小时微血管无扩张，但其周围水肿增加，24小时两者均改善。电镜下2、24小时毛细血管周水肿明显；胶质细胞胞体肿胀在2小时明显，24小时无显著差异；NF在2小时能减轻胶质细胞肿胀程度，但对毛细血管周水肿范围影响不大。结论 形态计量学方法能准确反映脑外伤后微循环及脑水肿改变。外伤早期慎用血管扩张药物。     

Chronic cerebrovascular dysfunction after traumatic brain injury

Traumatic brain injuries (TBI) often involve vascular dysfunction that leads to long‐term alterations in physiological and cognitive functions of the brain. Indeed, all the cells that form blood vessels and that are involved in maintaining their proper function can be altered by TBI. This Review focuses on the different types of cerebrovascular dysfunction that occur after TBI, including cerebral blood flow alterations, autoregulation impairments, subarachnoid hemorrhage, vasospasms, blood–brain barrier disruption, and edema formation. We also discuss the mechanisms that mediate these dysfunctions, focusing on the cellular components of cerebral blood vessels (endothelial cells, smooth muscle cells, astrocytes, pericytes, perivascular nerves) and their known and potential roles in the secondary injury cascade. © 2016 Wiley Periodicals, Inc.     

Inhibition of nitric oxide synthase aggravates brain injury in diabetic rats with traumatic brain injury

Studies have shown that hyperglycemia aggravates brain damage by affecting vascular endothelial function. However, the precise mechanism remains unclear. Male Sprague-Dawley rat models of diabetes were established by a high-fat diet combined with an intraperitoneal injection of streptozotocin. Rat models of traumatic brain injury were established using the fluid percussion method. Compared with traumatic brain injury rats without diabetic, diabetic rats with traumatic brain injury exhibited more severe brain injury, manifested as increased brain water content and blood-brain barrier permeability, the upregulation of heme oxygenase-1, myeloperoxidase, and Bax, the downregulation of occludin, zona-occludens 1, and Bcl-2 in the penumbra, and reduced modified neurological severity scores. The intraperitoneal injection of a nitric oxide synthase inhibitor N(5)-(1-iminoethyl)-L-ornithine(10 mg/kg) 15 minutes before brain injury aggravated the injury. These findings suggested that nitric oxide synthase plays an important role in the maintenance of cerebral microcirculation, including anti-inflammatory, anti-oxidative stress, and anti-apoptotic activities in diabetic rats with traumatic brain injury. The experimental protocols were approved by the Institutional Animal Care Committee of Harbin Medical University, China(approval No. ky2017-126) on March 6, 2017.     

Assessment of regional blood flow in cerebral motor and sensory areas in patients with spinal cord injury

We assessed the presence and the degree of alteration of the regional blood flow (rCBF) as visualized by Tc-99m HMPAO brain rest SPECT in the sensory motor cortex and subcortical structure in spinal cord injury (SCI) patients, who suffered from various levels of motor and sensory function loss. Twenty-two patients (mean age: 42.1+/-13.4 years, 18 M, 4 F) and 11 control subjects (mean age: 32.2+/-6.4 years, 8 M, 3 F) participated in this study. The spinal cord injury group was consisted of 2 groups (14 paraplegic and 8 tetraplegic patients). The corticocortical rCBF ratios were calculated by using region of interests obtained from 34 cortical areas on coronal slices. Significantly reduced rCBF were measured from 11 cortical areas in tetraplegic patients and 11 cortical areas in paraplegic patients. Some of these areas were different in each group. In the tetraplegic group, significant reduction was observed in the following rCBF areas: left anterior cingulate gyrus, left medial supplementary motor area, bilateral front and back aspects of posterior cingulate gyrus, right lateral primary motor area, right medial primary sensory area, bilateral putamen, and right cerebellum. In the paraplegic group, reduced rCBF areas were as follows: bilateral anterior cingulate gyrus, right lateral supplementary motor area, left front aspect of posterior cingulate gyrus, left lateral primary motor area, bilateral back aspects of posterior cingulate gyrus, right medial primary sensory area, left lateral primary sensory area and bilateral putamen. In conclusion, in some of the movement-cortical and subcortical areas having significantly reduced blood flow in SCI may be helpful to demonstrate the disrupted areas of rCBF by SPECT. We believe that it may be useful if these findings should be considered during the evaluations related to the reorganization in SCI cases.     

依达拉奉对脑缺血再灌注大鼠血管内皮生长因子和血管生成素-1的影响

目的探索依达拉奉对脑缺血再灌注大鼠的血管内皮生长因子(VEGF)和血管生成素-1(Ang-1)的影响。方法采用大脑中动脉闭塞法建立大鼠脑缺血再灌注模型,将雄性SD大鼠随机分为3组:假手术组、模型组和依达拉奉组,每组19只。HE染色观察梗死灶周围的病理改变,透射电镜下观察梗死灶周围脑组织的毛细血管内皮形态变化,激光多普勒微循环血流仪监测微循环血流量,采用免疫组化检测脑组织VEGF和Ang-1的表达。结果依达拉奉组的组织病理损伤和血管内皮细胞损伤要轻于模型组,微循环血流量(47.32±6.58)明显高于模型组(40.51±7.96)(P0.05)。模型组大脑组织的VEGF和Ang-1[(0.124±0.021),(0.099±0.014)]均明显高于假手术组[(0.118±0.018),(0.095±0.016)](均P0.05),而依达拉奉组大脑组织的VEGF和Ang-1[(0.147±0.019),(0.124±0.021)]均明显高于模型组[(0.136±0.023),(0.118±0.018)](均P0.05)。结论依达拉奉能上调缺血再灌注大鼠脑组织的VEGF和Ang-1来保护梗死灶周围毛细血管的内皮细胞。     

颅骨成形术对脑血流灌注的影响：SPECT—CT显像观察

目的采用SPECT—CT研究颅骨成形术对脑血流灌注的影响。方法19例重型颅脑损伤病人在标准大骨瓣减压术后3个月行颅骨成形术。术前2d及术后10d行SPECT—CT检查,SPECT—CT检查当天及术后1个月行神经功能评价,分析手术前后伤侧及对侧海马、皮质、丘脑的脑血流灌注值。结果颅骨成形术前后病人神经功能评价差异无统计学意义。SPECT—CT脑血流灌注显像提示对侧运动感觉皮质、海马、丘脑脑血流量在颅骨成形术前后未见显著差异。伤侧运动感觉皮质脑血流量在颅骨成形术后明显增加（P〈0．05）,与对侧比较无显著差异。伤侧丘脑脑血流量在颅骨成形术后明显增加（P〈0．05）,但仍低于对侧。伤侧海马脑血流量在颅骨成形术后无显著变化,但低于对侧检测值。结论颅骨成形术可显著改善伤侧皮质及丘脑血流灌注状况。     

Capillary transit time heterogeneity and flow-metabolism coupling after traumatic brain injury

Most patients who die after traumatic brain injury (TBI) show evidence of ischemic brain damage. Nevertheless, it has proven difficult to demonstrate cerebral ischemia in TBI patients. After TBI, both global and localized changes in cerebral blood flow (CBF) are observed, depending on the extent of diffuse brain swelling and the size and location of contusions and hematoma. These changes vary considerably over time, with most TBI patients showing reduced CBF during the first 12 hours after injury, then hyperperfusion, and in some patients vasospasms before CBF eventually normalizes. This apparent neurovascular uncoupling has been ascribed to mitochondrial dysfunction, hindered oxygen diffusion into tissue, or microthrombosis. Capillary compression by astrocytic endfeet swelling is observed in biopsies acquired from TBI patients. In animal models, elevated intracranial pressure compresses capillaries, causing redistribution of capillary flows into patterns argued to cause functional shunting of oxygenated blood through the capillary bed. We used a biophysical model of oxygen transport in tissue to examine how capillary flow disturbances may contribute to the profound changes in CBF after TBI. The analysis suggests that elevated capillary transit time heterogeneity can cause critical reductions in oxygen availability in the absence of ‘classic'' ischemia. We discuss diagnostic and therapeutic consequences of these predictions.