大鼠坐骨神经切断后腰脊髓腹角内胶质细胞和神经元的可塑性变化

目的：探讨大鼠单侧坐骨神经切断后腰脊髓腹角胶质细胞和运动神经元的反应及其相互关系。方法：用免疫组织化学技术、HE染色和Tunnel法，观察坐骨神经切断后1，6，12，24h及3，7和14d腰脊髓腹角胶质原纤维酸性蛋白(GFAP)标记的星形胶质细胞、OX-42标记的小胶质细胞及运动神经元的变化。结果：坐骨神经切断侧腰脊髓腹角可见星形胶质细胞和小胶质细胞活化，星形胶质细胞的活化早于小胶质细胞；后期运动神经元发生凋亡，HE染色显示凋亡细胞周围为反应性OX-42阳性小胶质细胞和GFAP阳性星形胶质细胞包绕。结论：研究结果提示，坐骨神经切断后切断侧腰脊髓腹角活化的胶质细胞与凋亡的运动神经元之间关系密切。     

间歇性θ爆发式磁刺激对高血压大鼠胼胝体髓鞘及局部炎症反应的影响

目的探究间歇性θ爆发式磁刺激对慢性高血压大鼠胼胝体区域髓鞘脱失、星形胶质细胞增生和小胶质细胞活化的改善作用。方法对雄性Sprague-Dawley大鼠随机进行双肾双夹术,制作易卒中型肾血管性高血压大鼠模型。术后22周,模型制备成功的高血压大鼠随机接受连续14 d间歇性θ爆发式磁刺激治疗(intermittent theta burst stimulation,iTBS)(高血压iTBS组,n=6)或假性刺激(高血压假刺激组,n=6),假手术大鼠接受假性刺激(假手术假刺激组,n=6)。HE染色观察胼胝体小动脉形态。免疫荧光染色观察胼胝体MBP标记的髓鞘脱失情况。免疫荧光染色观察GFAP标记的星形胶质细胞数量和IBa-1标记的小胶质细胞数量和形态,以评价星形胶质细胞增生和小胶质细胞活化。结果高血压iTBS组和高血压假刺激组出现明显小动脉管壁增厚。与假手术假刺激组相比,高血压假刺激组胼胝体MBP阳性面积比例减少(P<0.01),GFAP阳性细胞和IBa-1阳性细胞数量明显增加(P<0.01),IBa-1阳性细胞胞体增大,突起变粗,分枝变少。iTBS治疗明显增加了高血压大鼠胼胝体MBP阳性面积比例,降低GFAP阳性细胞和IBa-1阳性细胞数量(P<0.01),IBa-1阳性细胞胞体变小,突起变细,分枝变多。结论iTBS治疗可减轻高血压大鼠胼胝体区域髓鞘脱失,抑制星形胶质细胞增生和小胶质细胞活化。     

睫状神经营养因子对坐骨神经切断吻合后大鼠脊髓前角胶质纤维酸性蛋白

背景：睫状神经营养因子具有多种生物活性,在神经系统发育、分化和损伤修复中具有重要意义。 目的：观察睫状神经营养因子对坐骨神经切断吻合后大鼠相应脊髓节段前角星形胶质细胞的特异标记物胶质纤维酸性蛋白表达的影响。 方法：将SD大鼠随机分为对照组、模型组、生理盐水组及药物组。除对照组外,对所有大鼠实施双侧坐骨神经切断吻合术,药物组手术区局部注射睫状神经营养因子100 ng/kg,1次/d,生理盐水组局部注射等量生理盐水。术后1,3,7,14,21,28 d取相应脊髓节段,免疫组织化学染色观察胶质纤维酸性蛋白的表达,苏木精-伊红染色、TUNEL染色对脊髓前角神经元进行计数。 结果与结论：大鼠坐骨神经切断吻合后相应脊髓节段星形胶质细胞胞体大,突起分枝多且粗大,神经元数目逐渐减少,凋亡神经元增多,胶质纤维酸性蛋白表达增高。与模型组和生理盐水组比较,药物组神经元存活数目增多,凋亡减少,胶质纤维酸性蛋白表达明显增加(P < 0.05或P < 0.01)。同时,药物组大鼠的运动功能障碍较轻,恢复较快。说明睫状神经营养因子可以通过促进大鼠脊髓前角胶质纤维酸性蛋白的表达起到神经保护作用。 关键词：胶质纤维酸性蛋白;睫状神经营养因子;星形胶质细胞;神经元凋亡;周围神经损伤     

氯胺酮对NMDA诱导大鼠脊髓背角星形胶质细胞损伤的保护作用

目的观察氯胺酮对N-甲基-D-天冬氨酸(NMDA)受体过度激活诱导大鼠脊髓背角星形胶质细胞凋亡影响,并探讨其可能的作用机制.方法取新生2～3 d wistar大鼠T11-L6脊髓背角星形胶质细胞原代纯化培养,GFAP鉴定星形胶质细胞纯度达98%后用于实验.将细胞随机分6组:对照组(C组),NMDA组(N组),氯胺酮组(K组)和三种不同浓度氯胺酮加NMDA组(0.1,0.5,1 mmol/L,标记为NK1～NK3组),再培养24 h后检测SOD活性和MDA含量,免疫组化HE复染观察Bcl-2蛋白和形态学变化,流式细胞仪检测星形胶质细胞凋亡率.结果N组细胞发生了大量凋亡,SOD活性显著降低,MDA含量明显增加,Bcl-2蛋白表达不明显;NK3组细胞凋亡被显著抑制,Bcl-2蛋白强阳性表达,SOD活性明显增加和MDA含量低.结论NMDA受体过度激活可诱导大鼠脊髓背角星形胶质细胞大量凋亡,适量氯胺酮显著抑制了细胞凋亡,其机制可能是增强了星形胶质细胞Bcl-2蛋白表达,同时抑制了自由基的产生和增强了SOD活性.     

缺血缺氧对体外培养星形胶质细胞细胞周期和增殖的影响

目的 观察缺血缺氧损伤对星形胶质细胞细胞周期和增殖的影响。方法 用流式细胞仪检测缺血缺氧后不同时问点星形胶质细胞细胞周期变化，并用荧光免疫细胞化学技术测定胶质细胞纤维酸性蛋白(GFAP)和增殖细胞核抗原(PCNA)的表达水平。结果 体外缺血缺氧损伤后星形胶质细胞S期较正常组明显增高，6h达高峰，而随后则呈下降趋势。PCNA阳性反应损伤后表达均增加，6h表达最高；在缺血缺氧早期，GFAP阳性染色增强，6h最高；缺血缺氧12h后GFAP阳性染色变弱。结论 缺血缺氧损伤后星形胶质细胞活化进入增殖期；PCNA参与了损伤后星形胶质细胞的修复和增殖；细胞周期事件与星形胶质细胞的活化密切相关。     

星形胶质细胞和神经元凋亡的细胞周期时相分布特征

目的 通过测定星形胶质细胞和神经元各自细胞凋亡的细胞周期特异性分布规律,以进一步探讨两种细胞不同的凋亡发生机制. 方法 建立大脑中动脉阻塞再灌注模型并取损伤后3 d的脑组织进行分析,缺血再灌注3 d后取脑,采用BrdU方法标记S期的细胞;再利用流式细胞分选技术特异性的将星形胶质细胞和神经元分选出来,采用BrdU标记并结合TUNEL方法,分析星形胶质细胞和神经元凋亡的细胞周期定位. 结果 与正常组比较,缺血再灌注组中缺血侧大脑皮层可见BrdU标记阳性细胞和TUNEL标记阳性细胞均明显增多,并可见TUNEL阳性和BrdU阳性双重标记的细胞.星形胶质细胞凋亡可以发生在细胞周期的各个时相,大多发生在G0/G1期,比神经元在G0/G1期的细胞凋亡率显著增高(P＜0.01);神经元进入S期的细胞凋亡率显著高于星形胶质细胞(P＜0.01). 结论 星形胶质细胞和神经元的细胞凋亡是多点启动的,脑缺血后两种细胞的凋亡具有不一致的细胞周期时相性分布特征.     

胶质纤维酸性蛋白与中枢神经系统疾病

<正>胶质纤维酸性蛋白(Glial Fibrillary Acidic Protein,GFAP) 是分子量为50-52kD的酸性蛋白,是星形胶质细胞的主要成分 之一,富含谷氨酸和天冬氨酸,以中间微丝蛋白和可溶性蛋白 两种形式存在于胶质细胞的胞浆中,是星形胶质细胞的骨架蛋 白。GFAP在星形胶质细胞受到刺激引起反应时,其表达发生 变化,因此GFAP能够用来特异性地标记星形胶质细胞、并被 认为是星形胶质细胞活化的标志。活化的星形胶质细胞 内GFAP的增高,可能起保护神经元的作用;星形胶质细胞在 损伤区分泌多种神经营养因子,以促进中枢神经和周围神经轴     

X线照射后体外培养星形胶质细胞VEGF和GFAP表达变化及意义

目的 研究X线照射体外培养的星形胶质细胞后血管内皮生长因子(VEGF)和胶质原纤维酸性蛋白(GFAP)表达随时间及剂量的变化,探讨星形胶质细胞与放射性脑损伤(RBI)的关系. 方法 以5、10、15、20 Gy剂量的X线照射体外原代培养的星形胶质细胞后继续培养48 h,或以20 Gy剂量照射后分别培养4、12、24、48 h,实验均设正常对照组即未照射组.免疫荧光染色检测GFAP观察各组细胞形态学的变化;DAPI染色观察星形胶质细胞的凋亡;Western blotting检测细胞GFAP、VEGF蛋白的表达情况. 结果 与正常对照组比较,不同剂量照射组星形胶质细胞增生、增多,变形,胞体肥大肿胀,分支增多,突起增粗,GFAP染色加深,且这种变化随照射剂量和时间增加而表现更明显;与正常对照组相比,各剂量照射组星形胶质细胞凋亡率差异无统计学差异(P＞0.05); Western blotting检测显示不同剂量组、20Gy剂量照射不同时间组星形胶质细胞GFAP、VEGF蛋白的表达均不同,差异有统计学意义(P＜0.05).与正常对照组相比,5、10、15、20 Gy照射组和20 Gy剂量照射后各时间组GFAP、VEGF的蛋白表达均增高,且随着照射剂量的增加GFAP、VEGF的蛋白表达亦增高,差异有统计学意义(P＜0.05).20 Gy剂量照射后4～48 h内GFAP的表达呈时间依赖性,20 Gy剂量照射后4～24 h内VEGF的表达呈时间依赖性. 结论 X线能诱导体外培养的星形胶质细胞活化,GFAP及VEGF的表达呈时间及剂量依赖性增高,VEGF异常高表达可能是造成RBI的重要原因.     

重复经颅磁刺激对大鼠海马、齿状回胶质纤维酸性蛋白、Ⅲ型补体受体表达的影响

目的观察重复经颅磁刺激(rTMS)对大鼠海屿、齿状间星形胶质细胞和小胶质细胞标记物胶质纤维酸性蛋白(GFAP)、Ⅲ型补体受体(OX-42)表达的影响。方法每日1次给予大鼠1Hz、100mT的TMS 10 min,共14d,观察大鼠行为的变化;14d后采用免疫组化ABC法检测其海马、齿状间GFAP、OX-42的表达,并与对照组比较。结果rTMS组、对照组大鼠海马、齿状MGFAP、OX-42阳性表达物在形态、数量等方面差异无显著性。结论本实验条件下的rTMS不会造成大鼠中枢神经系统明显损伤。     

大鼠下丘脑室旁核小胶质细胞对次声的反应

目的探讨次声作用后大鼠下丘脑室旁核小胶质细胞与星形胶质细胞的变化及其相互关系。方法将SD大鼠反复暴露于声压级16Hz130dB的次声环境中。用抗大鼠Ⅲ型补体受体标志物（0X42）和抗胶质细胞原纤维酸性蛋白（GFAP）的免疫组化方法，观察次声作用后即刻，7d，14d大鼠下丘脑室旁核小胶质细胞与星形胶质细胞的变化及其相互关系。结果正常大鼠下丘脑室旁核的小胶质细胞和星形胶质细胞的数量较少，一般为静息性形态，胞体小，突起细长，染色浅淡。次声作用后大鼠下丘脑小胶质细胞被激活，胞体变大，突起短粗，染色深，7d以后逐渐减弱；次声作用后第7d起星形胶质细胞变多，胞体变大，突起变粗，染色深，第14d达到高潮；小胶质细胞和星形胶质细胞之间的关系密切。结论次声作用后小胶质细胞比星形胶质细胞早被激活；两者的关系密切。     

Mechanisms of the suppression of the nociceptive reflex of the opening of the mouth during stimulation of the structures of the limbic brain

The comparative effectiveness of the inhibitory influence of tetanic stimulation of hypothalamus, amygdala and limbic cortex on EMG-response of m. digastricus evoked by electrical stimulation of tooth pulp nociceptive afferents was studied in cats anesthetized with a mixture of chloralose and nembutal. It was found that inhibition of the EMG-component of the jaw-opening reflex is most pronounced in case of stimulation of medial and lateral region of the hypothalamus, the inhibitory effect of central and medial nuclei of the amygdala is less pronounced and the effect of the limbic cortex is the weakest. It was shown that the mechanism of the antinociceptive effect of tetanic stimulation of the hypothalamus is not related to the concomitant increase of the blood pressure. After stabilization of the blood pressure the suppressive effect of the hypothalamus remains without changes, that points out to a direct, primary, not baro-afferent mechanism of the inhibition of the activity of nociceptive neurons of the trigeminal sensory nuclei. Noradrenaline, injected intravenously, induced a large increase of the blood pressure accompanied by a pronounced inhibition of the pain reflex. Angiotensin causes the same degree of blood pressure elevation without changes in the amplitude of the EMG-response of the pain reflex. Hypothalamic and noradrenergic mechanisms for control of pain sensitivity are discussed.     

药物治疗与合并认知行为治疗对强迫症疗效的比较

目的探讨认知行为心理治疗(CBT)在强迫症(OCD)患者各亚型治疗中的有效性和规律性。方法本研究为临床对照研究。符合入组标准的强迫症患者按患者自愿原则分为两组,治疗观察3、6、12个月。疗效评定分别运用Yale-Brown强迫量表,自拟的自评好转程度量表和临床疗效评定。结果认知行为心理治疗合并药物治疗组31例,临床有效率70.9%,其中治愈率1.8%。单纯药物治疗组24例,临床有效率33.3%。Yale-Brown强迫量表和自评量表得分在6个月和12个月两组有显著差异(P<0.05)。其中强迫症亚型(怕脏型、反复检查型和反复担心型)的疗效比较,怕脏型在治疗3个月末两组间自评量表评分有显著性差异(P<0.05);反复担心型在治疗6个月末两组间Yale-Brown强迫量表总分有显著性差异(P<0.05);反复检查型两组间无统计学差异。结论认知行为心理治疗合并药物治疗强迫症的疗效明显优于单纯药物治疗。强迫症的亚型在治疗中的有效性次序为:反复担心型>怕脏型>反复检查型。     

Characteristics of the sympathetic innervation of the nictitating membrane and of the vasculature of the nose and tongue of the cat

Summary Vasomotor responses from the nasal mucosa and tongue, and contractions of the nictitating membrane, were recorded on stimulation of the cervical sympathetic or internal carotid nerves.Preganglionic sympathetic nerve fibres which elicited a membrane response possessed a lower threshold than those which evoked nasal vasoconstriction, while the latter displayed a lower threshold than fibres which evoked tongue vasoconstriction. The sympathetic vasodilator fibres to the tongue, whose activity was revealed after-receptor blockade, had a similar threshold to the vasoconstrictor fibres.Membrane contraction, nasal vasoconstriction and occasionally tongue vasoconstriction could be evoked by stimulating the internal carotid nerve. The postganglionic fibres innervating the nasal mucosa had a similar threshold to those of the nictitating membrane, which may indicate that there are small myelinated fibres innervating the mucosa.The preganglionic compound nerve action potential had four major components, S₁–S₄. S₁, S₂ and usually S₃ fibres were associated with membrane contraction; S₂, S₃ and sometimes S₁ fibres were associated with nasal vasoconstriction; and S₃, usually S₂ and occasionally S₁ fibres were associated with vasoconstriction in the tongue. It is concluded that each of these three groups of nerve fibres, but not S₄ fibres, may include fibres associated functionally with the three effectors.There was a considerable difference between the relative amplitude of the responses of the three effectors elicited by stimulation of the cervical sympathetic nerve at frequencies between 0.2 and 2 Hz. Vasoconstrictor responses were relatively larger than membrane contractions suggesting differences in the mechanisms of neurotransmission at the neuroeffector junctions.     

Development of the projection from the nucleus of the brachium of the inferior colliculus to the superior colliculus in the ferret

Neurons in the deeper layers of the superior colliculus (SC) have spatially tuned receptive fields that are arranged to form a map of auditory space. The spatial tuning of these neurons emerges gradually in an experience-dependent manner after the onset of hearing, but the relative contributions of peripheral and central factors in this process of maturation are unknown. We have studied the postnatal development of the projection to the ferret SC from the nucleus of the brachium of the inferior colliculus (nBIC), its main source of auditory input, to determine whether the emergence of auditory map topography can be attributed to anatomical rewiring of this projection. The pattern of retrograde labeling produced by injections of fluorescent microspheres in the SC on postnatal day (P) 0 and just after the age of hearing onset (P29), showed that the nBIC-SC projection is topographically organized in the rostrocaudal axis, along which sound azimuth is represented, from birth. Injections of biotinylated dextran amine-fluorescein into the nBIC at different ages (P30, 60, and 90) labeled axons with numerous terminals and en passant boutons throughout the deeper layers of the SC. This labeling covered the entire mediolateral extent of the SC, but, in keeping with the pattern of retrograde labeling following microsphere injections in the SC, was more restricted rostrocaudally. No systematic changes were observed with age. The stability of the nBIC-SC projection over this period suggests that developmental changes in auditory spatial tuning involve other processes, rather than a gross refinement of the projection from the nBIC.     

A contribution to the study of the architecture of the autonomic nervous system of the digestive tract of the rat

Summary The distribution of aminergic and non-aminergic nerve fibres to the different constituents of the wall of the digestive tract in various regions is described. Aminergic fibres synapse with all nervous perikarya. Densely interlacing networks of nerve fibres are found in both layers of the tunica muscularis and in the lamina muscularis mucosae. A finely meshed plexus is observed in relation to the wall of the blood vessels in the wall of the gut. There are many fibres connecting the muscular and the vascular plexus. No nerve fibres have been observed in direct relation to the epithelium.The functional implications of these findings are discussed.