丹参在缺血再灌注时的神经保护作用——成纤维细…

丹参作为传统中药治疗缺血性脑血管病具有良好的疗效，本实验研究丹参是否通过影响成纤维细胞生长因子起修复作用，用大鼠线栓法缺血再灌注模型及免疫组化方法，发现大鼠缺血９０ｍｉｎ，再灌流４８ｈ后，缺血侧皮层，尾壳核及海马区ｂＦＧＦ样免疫反应增加及神经细胞变性；缺血后给予丹参大鼠，ｂＦＧＦ样免疫反应加强，并且缺血对应侧脑区也可见轻度的ｂＦＧＦ样免疫反应改变，且神经细胞变性程度较轻，提示丹参对脑缺血再灌流的保     

巴曲酶对局灶性脑缺血再灌流大鼠成纤维细胞生长因子样免疫反应的影响

以往实验研究曾发现巴曲酶对脑血管病具有良好的神经保护作用。本文研究的目的为：巴曲酶是否还通过影响成纤维细胞生长因子起修复作用。用大鼠中大脑动脉（ＭＣＡ）线栓法脑缺血再灌注模型及免疫组化方法发现在缺血９０ｍｉｎ，再灌流４８ｈ后，缺血侧皮层、尾壳核及海马区ｂＦＧＦ样免疫反应细胞增加及神经细胞变性。在缺血后给予巴曲酶（８ＢＵ／ｋｇ），ｂＦＧＦ样免疫反应加强，并且缺血对侧相应ＭＣＡ供血脑区也可见轻度的ｂＦＧＦ样免疫反应改变，神经细胞变性之程度较轻。提示：巴曲酶对脑缺血再灌注的保护作用可能与它加强ｂＦＧＦ的修复作用有关。     

巴曲酶对局灶性脑缺血再灌流大鼠成纤维细胞生长因子样免疫…

以往实验研究曾发现巴曲酶对脑血管病具有良好的神经保护作用，本文研究的目的为：巴曲酶是否还通过影响成纤维细胞生长因子起修复作用，用大鼠中大脑动脉（ＭＣＡ）线栓法脑缺血再灌注模型及免疫组化方法发现在缺血９０ｍｉｎ再灌流４８ｈ后，缺血侧皮层，尾壳核及海马区ｂＦＧＦ样免疫反应细胞增加及神经细胞变性，在缺血后给予巴曲酶（８ＢＵ／ｋｇ），ｂＦＧＦ样免疫反应加强，并且缺血对侧相应ＭＣＡ供血脑区也可见轻度的ｂＦＧ     

脑缺血再灌流后bFGF基因的表达及MK-801的影响

目的 研究脑缺血再灌流后ｂＦＧＦ基因表达的意义及其机制。方法 采用原位杂交和１免疫组化的方法,观察大鼠脑缺血再灌流后ｂＦＧＦ基因的表达及ＭＫ－８０１对它们的影响。结果 缺血２ｈ再灌流１ｈ可见ｂＦＧＦ表达增高（Ｐ〈０．０５）,ｂＦＧＦ ｍＲＮＡ表达于１２ｈ达高峰,ｂＦＧＦ蛋白于２４ｈ,达高峰;ＭＫ－８０１组与缺血组相比,于再灌流６ｈ～４８ｈ ｂＧＦＧ表达减弱（Ｐ〈０．０５）。结论 局灶性脑缺血再灌流     

大鼠脑缺血再灌流后HSP70的表达和bFGF对其影响

目的研究脑缺血再灌流后热休克蛋白７０（ＨＳＰ７０）表达的意义及外源性碱性成纤维细胞生长因子（ｂＦＧＦ）对其影响。方法应用免疫组化的方法观察大鼠局灶性脑缺血再灌流后脑组织ＨＳＰ７０的表达以及在侧脑室注射外源性ｂＦＧＦ对其影响。结果缺血２小时再灌流０小时即可见ＨＳＰ７０表达,至２４小时达高峰。ｂＦＧＦ组与生理盐水组相比于６～４８小时各组可见ＨＳＰ７０表达增高（Ｐ〈０．０５）。结论脑缺血诱导ＨＳＰ７０的     

实验性脑缺血及再灌注大鼠脑组织bFGF表达的研究

目的　探讨大鼠脑缺血２ ｈ 后不同时间再灌注脑组织碱性成纤维细胞生长因子（ｂＦＧＦ）的表达程度。方法　采用线栓法栓堵一侧大鼠大脑中动脉,建立脑缺血及再灌注模型。用免疫组化的方法,对缺血脑组织ｂＦＧＦ的表达进行观察。用统计学分析ｂＦＧＦ阳性细胞的数据。结果　正常脑组织中只有微量ｂＦＧＦ的表达;脑缺血２ ｈ 后再灌注０．５ ｈ 起,ｂＦＧＦ少量表达,为神经元表达;脑缺血２ ｈ 后再灌注２２ ｈ,ｂＦＧＦ表达达到高峰,神经元及神经胶质细胞均有表达。脑缺血２ ｈ 后再灌注１６６ ｈ,ｂＦＧＦ仍有持续表达。结论　ｂＦＧＦ参与缺血脑组织的修复过程     

局灶性缺血再灌注时丹参对热休克蛋白70的影响—免疫细胞化学及…

丹参素以活血化瘀称著，常用于治疗缺血性脑血管病，热休克蛋白７０（ＨＳＰ７０）具神经保护作用。本文研究目的：丹参是否也通过影响ＨＳＰ７０起作用，采用大鼠犬脑中动脉（ＭＣＡ）线检模型。以免疫细胞化学及病理组织方法在同一动物中进行研究。结果发现在缺血９０ｍｉｎ再灌注２４ｈ后，对照组手术侧皮层ＨＳＰ７０免疫反应增强及皮层神经细胞有缺血性损伤，而丹参组（缺血９０ｍｉｎ时给予丹参１０ｇ／Ｋｇ，体重ｉｐ），则Ｈ     

bFGF和EGF对体外培养新生大鼠海马神经元促生长作用的研究

ｂＦＧＦ和ＥＧＦ对体外培养新生大鼠海马神经元促生长作用的研究吴承远苏万东张洪华王建刚为了研究碱性成纤维细胞生长因子（ｂＦＧＦ）和表皮生长因子（ＥＧＦ）在中枢神经系统中的生物学作用，本实验采用神经细胞体外原代分离培养的方法，来观察不同浓度ｂＦＧＦ和ＥＧ...     

大鼠脑缺血再灌注损伤后脑组织STAT3变化的免疫组织化学研究

目的 探讨信号转导和转录激活子（ＳＴＡＴ）３在大鼠局灶性脑缺血再灌注损伤中的表达及其与缺血性神经细胞损伤的关系方法 用ＡＢＣ免疫组化方法观察大鼠局灶性脑缺血再灌注损伤后脑组织中的ＳＴＡＴ３蛋白免疫反应阳性细胞分布。结果 正常和假手术大鼠脑内以及脑缺血后的非缺血半球脑组织中未发现有ＳＴＡＴ３免疫反应阳性细胞,脑缺血再灌注损伤后１２小时在栓塞侧梗死区可见少量ＳＴＡＴ３免疫阳性细胞,２４小时后阳性细胞显著增多达高峰,在缺血侧纹状体和缺血皮质周边区表达最明显,１周后梗死周边区少数神经细胞仍有阳性表达。差异有显著意义（Ｐ〈０．０１）。结论 ＳＴＡＴ３活化及超量表达可能介导了缺血神经细胞信号转导过程,并参与了脑缺血神经细胞损伤与修复的病理生理过程。     

c—Jun,bFGF,HSP70在丹参改善颞叶缺血大鼠空间记忆障碍中的？…

目的：探讨在丹参改善单侧缺单侧颞叶缺血大鼠空间记忆障碍中ｃ－Ｊｕｎ，ｂＦＧＦ和ＨＳＰ７０的表达变化。方法：采用立体定向光化学方法选择性地导致大鼠左侧颞叶皮层缺血，术前３０分钟及术后第３天分别给丹组组大鼠腹腔注射丹参１０ｇ／ｋｇ，用Ｍｏｒｒｉｓ水迷宫及图像自动监视系统监测大鼠行为。然后取脑 理学及ｃ－Ｊｕｎ、ｂＦＧＦ和ＨＳＰ７０免疫组化分析。结果：经丹参治疗，颞叶缺生按时完成大鼠的空间记忆障碍得到显     

Increase in basic fibroblast growth factor-like immunoreactivity in rat brain after forebrain ischemia

Using immunohistochemical techniques, a study was conducted to determine whether basic fibroblast growth factor (bFGF) is generated as one of the 'self-repair' responses in rat brain following transient forebrain ischemia. In normal brain, slight bFGF-like immunoreactivity was observed. However, in rats exposed to 20 min of forebrain ischemia, intense bFGF-like immunoreactivity was observed in the CA1 subfield of the hippocampus and the caudate putamen, and marked activity was evident in the temporal cortex, corpus callosum and the CA4 subfield of the hippocampus. Marked neuronal degeneration was also observed in these brain regions following forebrain ischemia. These results suggest that induction of bFGF-like immunoreactivity may be related to the healing which follows brain ischemia.     

Basic FGF in astroglial, microglial, and neuronal cultures: characterization of binding sites and modulation of release by lymphokines and trophic factors.

The present study characterizes whether basic fibroblast growth factor (bFGF) is present and released from astroglia, microglia, and hippocampal neurons in vitro. For cell content, bFGF-like immunoreactivity (IR) of cell extracts was measured, whereas release was determined by assessing the levels of bFGF-like IR in media. In addition, the effects of lymphokines and trophic factors that are known to be released from these cells on bFGF release were examined. For all three cell types, bFGF-like IR in extracts of cell lysates was detectable. In addition, media content was highest in astroglial cultures and lowest in neuronal cultures. Although bFGF-like IR of neuronal and microglial media appeared to increase with time in culture, this was likely due to significant astroglial proliferation. Thus, notable levels of bFGF are released by astroglia in vitro. In astroglia, bFGF release was enhanced by interleukin-1 (IL-1), IL-6, and epidermal growth factor (EGF), but not by other lymphokines or NGF. In contrast, bFGF in microglial media was reduced by IL-3, EGF, and NGF, but slightly augmented by gamma-interferon (IFN); other lymphokines were ineffective. In addition, no effects were seen in the neuronal cultures. It is likely that the bFGF found in glial media interacts with bFGF receptors since in both glial and neuronal cell types, a single class of low-capacity (Bmax), high-affinity (Kd) bFGF binding sites was evident. The possibility that endogenous bFGF acts as an autocrine factor for astroglia was further supported by experiments that tested the mitogenic effects of exogenous bFGF on glial cells. bFGF significantly enhanced 3H-thymidine uptake into astroglial, but not microglial, cells in vitro. Thus, the present study demonstrates that a complex regulation of glial bFGF release by astroglia and microglia occurs in vitro. Moreover, the results are consistent with an autocrine role for bFGF in astroglial cultures.     

Basic FGF-like protein in the lower stato-acoustic system of the neonatal and adult rat.

Basic fibroblast growth factor (bFGF)-like localization was studied immunohistochemically in the lower auditory tract of neonatal and adult rats. During the neonatal period, bFGF-like immunoreactivity is present in the cytoplasm of inner hair cells, spiral ganglion cells, Scarpa's ganglion cells, in auditory brain stem nuclei and in vestibular nuclei. At the adult stage, bFGF-like protein is widely distributed in the auditory brain stem but was not found in the cochlea. These results suggest that bFGF could be implicated in the development as well as in the neuronal maintenance and plasticity of the auditory system.     

碱性成纤维细胞生长因子治疗猫急性脑梗死的作用机制研究

目的:观察碱性成纤维生长因子(bFGF)处理的缺血再灌注不同时程的猫脑组织中微管相关蛋白(MAP-2)和神经丝蛋白(NTP)的表达,探讨bFGF治疗缺血性脑损伤的可能作用机制。方法:健康家猫30只,随机分为生理盐水对照组和bFGF治疗组。采用左侧眼眶入路制作大脑中动脉缺血再灌注模型。于术前和再灌注24h、48h和7d,采用Philip的猫脑缺血神经功能评分标准进行神经功能缺损评分;应用免疫组织化学SP法检测缺血再灌注不同时程的脑组织MAP-2及NTP蛋白表达,进行免疫阳性细胞计数。结果:缺血再灌注48h后,治疗组动物神经功能受损程度较对照组明显减轻,MAP-2及NTP蛋白阳性细胞数目较对照组也显著增加。结论:bFGF通过诱导MAP-2及NTP蛋白的表达,减轻了缺血再灌注脑组织的神经元损伤和促进了神经纤维生长,从而改善受损的神经功能。     

Fibroblast growth factor and hydrocephalus

The immunohistochemical localization of basic fibroblast growth factor (bFGF) was studied in ventricular ependyma and choroid plexus of aged-matched normotensive and spontaneously hypertensive rats at different ages using a polyclonal antibody against bFGF. The bFGF-like immunoreactivity was observed in brain ependyma and choroid plexus of young and old normotensive rats. However, a progressive loss of immunoreactivity was observed with age in spontaneously hypertensive rats, that was associated with a progressive cerebroventricular dilation. These results show a new neuroendocrine anomaly to be added to the many others previously observed in this rat strain, when they develop hydrocephalus as they age.     

Localization of basic fibroblast growth factor-like immunoreactivity in the rat brain.

The immunohistochemical localization of basic fibroblast growth factor (bFGF) was studied in the adult rat brain, using a specific antibody against a synthetic bFGF fragment (the N-terminal 12 residues). Widespread but uneven regional localization of bFGF-like immunoreactive neurons and fibers was observed. Ependymal cells were also stained. The immunoreactive neurons were found in the cerebral cortex, olfactory bulb, septum, basal magnocellular nuclei, thalamus, hypothalamus, globus pallidus, hippocampus, amygdala, red nucleus, central gray of the midbrain, cerebellum, dorsal tegmental area, reticular formation, cranial motor nuclei and spinal cord. Immunoreactive fiber bundles and nerve terminals were also detected. These results indicate that bFGF is produced by or present in a specific neuronal cell population of the central nervous system.     

血管内皮生长因子和碱性成纤维细胞生长因子在缺血预处理大鼠局灶性脑缺血再灌注区域的表达

背景：脑缺血预处理可增加碱性成纤维细胞生长因子的表达,可能导致脑缺血耐受的产生。大鼠大脑中动脉缺血再灌注给予血管内皮生长因子能够起到神经保护作用。 目的：观察缺血预处理对缺血再灌注大鼠血管内皮生长因子和碱性成纤维细胞生长因子表达的影响。 方法：将SD大鼠随机分为缺血预处理组、模型组和假手术组。缺血预处理及模型组线栓法阻塞大脑中动脉制备脑缺血模型。预处理组在脑缺血-再灌注前3 d用插入尼龙线阻塞大脑中动脉,缺血2 h后再灌注22 h。模型组第一次手术将线栓前推5 mm,不阻断血流,其他同预处理组。假手术组仅插入尼龙线不阻塞大脑中动脉。用苏木精-伊红染色法观察3组间神经细胞变化。用抗生物素-生物素-过氧化物酶复合物法检测各组血管内皮生长因子和碱性成纤维细胞生长因子蛋白的表达。分别比较3组神经功能评分、光镜下脑缺血再灌注区神经细胞形态、血管内皮生长因子和碱性成纤维细胞生长因子的表达。 结果与结论：与模型组比较,预处理组神经功能评分明显低于模型组(P < 0.01)。光镜下观察结果显示,与模型组比较,预处理组缺血面积及缺血程度均减轻,血管内皮生长因子和碱性成纤维细胞生长因子表达均明显升高(P < 0.05)。结果提示缺血预处理可能通过增强血管内皮生长因子和碱性成纤维细胞生长因子而对缺血再灌注大鼠神经细胞起保护作用。     

Induction of Tie-1 and Tie-2 receptor protein expression after cerebral ischemia-reperfusion.

Tie-1 and Tie-2 are receptor tyrosine kinases (RTKs) that are exclusively expressed in endothelial cells and play important roles in endothelial cell biology. The authors have reported previously the temporal profiles of Tie-1 and Tie-2 mRNA expression after focal cerebral ischemia-reperfusion. In the current study, the localization of Tie-1/Tie-2 mRNA and proteins were further investigated in the same focal ischemia model. In situ hybridization showed that, after 60-minute ischemia and 72-hour reperfusion, both Tie-1 and Tie-2 mRNA appeared as capillary-like structures in the ischemic middle cerebral artery (MCA) cortex. Western blot analysis showed a biphasic expression of Tie-1 protein in the same region. The first peak, spanning the ischemic and early reperfusion period. was of low intensity and short-lived. The second peak was of greater intensity and spanning the period from 72 to 168 hours after reperfusion. Similarly, Tie-2 expression at the protein level also exhibited a biphasic pattern. Immunohistochemical studies, after 72 hours of reperfusion, showed that although Tie-1 and Tie-2 were detected within the ischemic cortex, they actually were expressed in different populations of endothelial cells in different regions. In agreement with the in situ hybridization study, Tie-1 immunoreactivity appeared as capillary-like structures in cortical layers 2 to 4. Similar capillary-like appearance of Tie-2 immunoreactivity was noted in the outer cortical layers. In addition, Tie-2 immunoreactivity also was observed in cortical layer 6b, where de novo large vessel formation was noted. Cellular colocalization experiments revealed that Tie-2 is expressed in proximity to its antagonist, Angpo-2, as well as basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in cortical layer 1, where active vessel remodeling was noted. Interestingly, bFGF only partially colocalized with VEGF, suggesting differential roles for these angiogenic factors during vessel remodeling. Tie-1 protein, to a lesser degree, also colocalized with Angpo-2, bFGF, and VEGF in cortical layer 1. Magnetic resonance imaging (MRI) showed increased regional cerebral blood flow (CBF) corresponding to the expression of these angiogenesis gene products. Together, these findings suggest that the evolving expression of angiogenesis genes underlie the robust vascular remodeling after ischemia and reperfusion.     

Intravenous basic fibroblast growth factor (bFGF) decreases DNA fragmentation and prevents downregulation of Bcl-2 expression in the ischemic brain following middle cerebral artery occlusion in rats

In previous studies, we showed that basic fibroblast growth factor (bFGF) reduced infarct volume when infused intravenously in animal models of focal cerebral ischemia. In the current study, we examined the potential mechanism of infarct reduction by bFGF, especially effects on apoptosis within the ischemic brain. We found that bFGF decreased DNA fragmentation in the ischemic hemisphere, as assessed by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) histochemical methods combined with morphological criteria. bFGF also prevented reduction of immunoreactivity of the anti-apoptotic protein Bcl-2 in the ischemic hemisphere, but did not alter immunoreactivity of the pro-apoptotic proteins Bax, Caspase-1, or Caspase-3. These changes in TUNEL histochemistry and Bcl-2 immunoreactivity were especially prominent in cortex at the borders ('penumbra') of infarcts, spared by bFGF treatment. We conclude that the infarct-reducing effects of bFGF may be due, in part, to prevention of downregulation of Bcl-2 expression and decreased apoptosis in the ischemic brain.