bFGF对大鼠脊髓损伤后神经细胞凋亡的影响

目的 :探讨大鼠脊髓损伤后应用碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor ,bFGF)对脊髓损伤区细胞凋亡的影响。方法 :利用Allen氏WD(weightdrop ,WD)技术 ,以 10 g× 2 .5cm致伤力造成SD大白鼠T8脊髓损伤模型 ,并于损伤平面以下蛛网膜下腔置细塑料导管。bFGF治疗组 (A组 )分别于术后即刻 1、2、4、8、12、2 4及 48h经导管注入bFGF溶液 2 0 μl(含bFGF10 0 u) ,以后每周经导管注入 2 0 μlbFGF ;对照组 (B组 )则在同时间注入等量生理盐水。损伤后 1、3、7、14、2 8d对脊髓损伤区进行细胞凋亡的检测 (TUNEL) ,采用计算机图像分析技术进行定量分析。结果 :A、B两组中均发现凋亡细胞 ,B组细胞凋亡率大于A组。结论 :bFGF能抑制脊髓损伤后脊髓损伤区的细胞凋亡。     

大鼠损伤脊髓提取液对骨髓基质细胞培养特性的影响

目的：探讨损伤脊髓提取液对骨髓基质细胞增殖和分化的影响。方法：将正常、伤后1周及伤后6周脊髓提取液加入细胞培养基中，以四唑盐法及免疫组化方法观察其对骨髓基质细胞增殖及抗原表达的影响。结果：加入脊髓提取液后对骨髓基质细胞的增殖无明显影响，但抗原表达发生变化，纤维连接蛋白(FN)表达减弱，且出现神经丝蛋白(NF)和胶质纤维酸性蛋白(GFAP)的表达，细胞形态出现神经细胞样改变。结论：骨髓基质细胞可能是一种脊髓损伤修复的理想细胞。     

实验性大鼠胚胎脊髓移植对损伤脊髓影响的观察

目的 建立了一个胚胎脊髓（ｆｅｔａｌ ｓｐｉｎａｌ ｃｏｒｄ）组织的低温保存方法笔一个损伤移植模式。方法 通过病理学、组织化学和免疫组织化学的研究,选用胎龄１４天的Ｗｉｓｔａｒ大鼠,分离胚胎脊髓,逐步降温,最后置于液氮中保存。１５天后取出,快速复温,并植入损伤大鼠脊椎内。全部动物于伤后８周处死。结果 经过低温保存的移植物能够在异体脊髓内存活并与宿主组织融合,其在损伤移植区的胶质细胞增生少于对照组。     

细胞外ATP促进体外培养大鼠脊髓前角运动神经元存活的研究

目的探讨细胞外ATP对体外培养的乳鼠脊髓前角运动神经元的作用。方法将ATP加入培养乳鼠前角运动神经元的培养基中,用PBS培养作为对照组。接种后,采用相差倒置显微镜观察细胞生长情况，并以计数绘制细胞生长曲线；用MTT比色法测定培养细胞的存活率。结果实验组细胞的存活率和轴突的长度明显优于对照组(t=7.157、5.784、7.186，P<0.01),实验组内各时间组比较，差异无显著性意义（t=0.639、1.72、0.804,P>0.O5)。结论细胞外给以ATP具有较强的维持神经元存活,促进轴突生长的作用。     

福尔马林致痛对大鼠行为学及脊髓NO的影响

目的：探讨大鼠福尔马林致痛模型脊髓背角一氧化氮合酶(NOS)的变化。方法：雄性SD大鼠64只，随机分为四组：生理盐水对照组(NS组)、福尔马林对照组(F组)、L-精氨酸组(LaF组)和L-NG-硝基精氨酸甲酯(LnF组)组。F组给予5％福尔马林100μl足底注射；LaF和LnF组在同F组处理前分别给予L-精氨酸(L-Arg)和L-NG-硝基精氨酸甲酯(L-NAME)。在福尔马林处理后30min、O～1h分别检测大鼠脊髓NOS的表达及观察大鼠的行为学表现。结果：F组的缩腿舔爪时间和脊髓的NOS表达显著长于、强于NS组；预先给予L-Arg或L-NAME分别能加强或抑制以上作用。结论：福尔马林致痛能引起大鼠脊髓背角一氧化氮(NO)的释放，可能是其产生伤害作用的机制之一。     

白细胞介素-10对大鼠脊髓损伤后神经细胞凋亡的影响

目的 观察白细胞介素-10(IL-10)对大鼠脊髓损伤(SCI)后脊髓细胞凋亡的影响,探讨IL-10对脊髓损伤的保护作用。方法 将SD大鼠随机分为3组:单纯SCI组(A组)、IL-10治疗组(B组)及假损伤组(Sham组)。除Sham组不致伤脊髓外,A、B两组应用改良的Allen打击法制作大鼠急性 SCI模型,B组大鼠于 SCI后 30 min腹腔注射 10 μg IL-10,再分别于伤后 12、24、48 h、4、8、16和 24 d应用苏木素-伊红(HE)染色、原位末端脱氧核糖核苷酸转移酶介导的脱氧尿苷三磷酸生物素缺口末端标记技术(TUNEL检测法)及开放野行为评分(BBB评分)观察IL-10治疗后脊髓细胞凋亡的变化及神经功能的恢复情况。结果 假损伤组未见凋亡细胞。A组大鼠伤后 12 h即可见零星的凋亡细胞,至 24 h渐增多,伤后 48 h脊髓组织凋亡细胞率达峰值。B组伤后 24、48 h及4d细胞凋亡率比单纯SCI组明显减少,差异具有显著性(P<0.01或P<0.05)。B组脊髓功能恢复亦比A组有明显提高(P<0.05)。结论 SCI后早期应用IL-10可抑制大鼠脊髓细胞凋亡,从而对脊髓损伤起到保护作用。     

大鼠脊髓背根神经节神经元的纯化培养

目的：建立一种简单、稳定、高效的大鼠脊髓背根神经节神经元原代纯化培养方法。方法：取新生SD大鼠的脊髓背根神经节，采用胰蛋白酶消化法，制成单细胞悬液，加入阿糖胞苷以纯化细胞，培养于含10％胎牛血清与重组人胶质细胞源性神经营养因子的DFI2培养基中，观察神经元生长状况，用细胞计数和神经元特异性烯醇化酶（NSE）免疫细胞化学染色测定细胞纯度。结果：培养的脊髓背根神经节神经元生长状态正常，有神经突起生长和标记蛋白的表达，可达到90％左有的纯度。结论：本培养方法简单、稳定、高效，为对神经元进一步的深入研究提供了实验模型．     

成年脊髓损伤大鼠脊髓神经干细胞的体外培养及分化研究

目的 探讨大鼠脊髓损伤后脊髓神经干细胞的分离培养方法及分化情况.方法 采用Allen法制作大鼠脊髓损伤模型,利用无血清培养和单细胞克隆技术在成年脊髓损伤7 d大鼠脊髓中分离具有单细胞克隆能力的神经干细胞,并进行培养鉴定.结果 从成年脊髓损伤7 d大鼠脊髓中成功分离出神经干细胞,该细胞具有连续克隆能力,可传代培养,表达神经巢蛋白抗原.分化后的细胞表达神经元细胞、星形胶质细胞和少突胶质细胞的特异性抗原.结论 致伤7 d的成年大鼠脊髓组织体外町培养出神经十细胞,并分化为神经无细胞、星形胶质细胞和少突胶质细胞,有可能参与脊髓损伤的修复过程.     

碱性成纤维细胞生长因子对体外培养胚胎大鼠脊髓神经元促生长活性的研究

目的：研究碱性成纤维细胞生长因子（basic fibroblast growth facfor,bFGF)对体外培养脊髓神经元的作用。方法：取E14大鼠脊髓神经组织,进行体外原代培养,分别加和不同浓度的bFGF。在相差显微镜下观察细胞生长发育情况,并测定其细胞内琥珀酸脱氢酶的活性。实验结果用t检验分析。结果：bFGF处理组神经元密度明显高于对照组,神经元胞体大而饱满,突起较长。细胞内琥珀酸脱氢酶活性较高,培养20d后神经元存活的数量也明显高于对照组。结论：bFGF能促进脊髓神经元在体外培养早期的存活,增加神经元的胞体直径和突起长度,增强神经元细胞内琥珀酸脱氢酶的活性,并能延缓体外培养脊髓神经元的衰老。     

脊髓损伤对大鼠骨转换及骨密度的影响

目的：探讨脊髓损伤（spinal cord injury,SCI）对大鼠骨转换及骨密度的影响。方法：60只3月龄SD大鼠均分为SCI组与对照组。SCI组于T10处完全横断脊髓;对照组仅行椎板切除术。术后1、3、6周时处死动物测血清钙（Ca）、磷（P）、碱性磷酸酶（ALP）、尿钙、磷、肌酊（Cr）以及股骨、股骨骨密度（bone mineral density,BMD）。:桂花 血钙、尿钙、尿钙／肌酝在伤后不同时间段均升高;血ALP在伤后1周时显著下降,3、6周时恢复正常。胫骨近端,股骨远、近端的BMD在6周时较对照组下降且差异显著。结论：SCI大鼠可见明显的骨转换增高以及破骨活性增强,其生化、骨密度的改变与人体有较好的相关性。SD大鼠可用于评价SCI后骨代谢改变,具可以作为SCI后骨质疏松的模型。     

脊髓半切伤神经细胞凋亡的实验研究

目的 探讨细胞凋亡在脊髓半切损伤发病机理中的作用。方法 采用原位末端标记法，对大鼠半切伤的不同时间的凋亡细胞数目和部位进行观察分析，并作正常对照。结果半切后的灰质和自质均有细胞凋亡，神经元凋亡后形成空泡，胶原细胞凋亡多见。细胞凋亡发生在损伤后12h，7d达高峰，至5周趋于正常。结论 大鼠脊髓半切伤后神经元和胶质细胞均发生凋亡，而胶质细胞凋亡常见。     

大鼠脊髓损伤后巢蛋白在脊髓组织中的表达

目的探讨大鼠脊髓损伤后巢蛋白(nestin)的表达规律及其意义。方法30只Wister成年大鼠,随机分为正常对照组(A组)、损伤组(B组)。采用Allen打击模型(25g·cm),在T10段造成急性脊髓损伤,于损伤后1d、3d、1周、4周、8周进行取材,对距离损伤中心5mm处脊髓进行nestin免疫组化检测。应用图像分析软件进行nestin阳性区域面积侧算。结果A组脊髓室管膜细胞只可见极少数细胞胞浆内nestin表达,白质中几乎无表达。B组中nestin于损伤后24h表达于室管膜以及软膜,灰质和白质亦有少量表达,1周达到高峰(P<0.05),4周明显下降,8周时很少或几乎无表达。结论脊髓组织的许多部位可能存在具有分化和更新潜能的祖细胞,脊髓损伤后这些细胞被激活,在功能恢复中可能发挥着重要的作用。     

The prevention of iatrogenic spinal cord injury utilizing the evoked spinal cord potential

Summary The evoked spinal cord potential elicited by direct stimulation of the cord has been used clinically to monitor cord function in the course of operations on the spine. The technique used allows measurement of a relatively large amplitude of potential, which is fairly stable against anaesthetics and related drugs, by means of a simple recording system and is sensitive enough to indicate cord damage. Continuous monitoring can easily be carried out. We have encountered no complications when using this method on 99 patients.

---

Résumé Le potentiel évoqué provoqué par la stimulation directe de la moelle épinière a été utilisé en clinique pour contrôler la fonction de la moelle lors des interventions sur le rachis. Cette technique permet de mesurer une assez grande amplitude de potentiel, qui est relativement stable à l'égard des anesthésiques et d'autres drogues de même type, grâce à un système simple d'enregistrement; il est suffisamment sensible pour détecter des altérations de la moelle. Une surveillance continue peut aisément être effectuée. Aucun incident n'a été rencontré chez 99 malades lors de l'utilisation de cette méthode.

     

强啡肽对大鼠行为学和脊髓组织学改变的影响及受体机制

目的探明强啡肽(Dyn)对脊髓的损伤效应及其受体机制。方法观测大鼠鞘内注射DynA(1-13)或联合注射Kappa阿片受体拮抗剂nor-BNI或兴奋性氨基酸(EAA)的NMDA受体拮抗剂MK-801后的运动功能和脊髓病理学变化。结果注射30nmolDyn组3d时,Tarlov运动功能评分下降,脊髓前角神经元数目减少,GFAP阳性神经胶质细胞数轻度增生。14d时,Tarlov运动功能评分未恢复,神经胶质细胞增生明显。而鞘内联合注射100nmol nor-BNI、100nmol MK-801后3d时与单纯Dyn组结果相似,14d时Tarlov运动功能评分明显恢复,脊髓前角神经元数目较Dyn组多,GFAP阳性神经胶质细胞增生不明显,nor-BNI组与MK-801组间比较差异不显著。结论Dyn鞘内注射可使大鼠运动功能、脊髓组织损害,而nor-BNI或MK-801有对抗其损害作用。Dyn的病理作用是通过Kappa阿片受体和EAA的NMDA受体两种途径介导的。     

The effects of spinal cord injury on psychogenic sexual arousal in males

PURPOSE: We determined if the degree of preservation of sensory function in the T11-L2 dermatomes could be used to determine the potential for psychogenic erectile responses in men with spinal cord injury. MATERIALS AND METHODS: Subjects included 45 men with spinal cord injury and 16 able-bodied control subjects. A 78-minute laboratory based analysis was done of subject subjective arousal, penile circumference, blood pressure, and heart rate responses to audiovisual erotic and audiovisual erotic combined with manual penile stimulation. RESULTS: Able-bodied subjects generally had significantly greater penile circumferences than spinal cord injured subjects during the stimulation periods. The degree of preservation of combined pinprick and light touch sensation in the T11-L2 dermatomes distinguished those who did and did not have a significant increase in penile circumference with audiovisual stimulation. Blood pressure and heart rate readings were generally higher in able-bodied than spinal cord injured subjects throughout the experimental protocol. However, all readings were within normal limits. CONCLUSIONS: Results support the hypothesis that psychogenic erection depends on the sympathetic nervous system. Findings underscore a possible parallel in neurological control of sexual responses between the sexes.     

神经节苷脂对大鼠脊髓损伤后微管相关蛋白-2表达的影响及意义

[目的]通过观察单唾液酸四己糖神经节苷脂(monosialotetrahexosyl gangliosides,GM-1)对于大鼠脊髓损伤(spinal cord injury,SCI)后微管相关蛋白-2(microtubule-associated protein 2,MAP-2)表达及运动功能恢复是否能够产生影响来探讨CM-1对大鼠脊髓损伤后神经细胞的保护作用及其机制.[方法]Wistar雌性大鼠66只(体重260～300 g),随机取6只作为正常对照组,余60只采用改良Allen's打击法于T9-11节段椎管制作大鼠急性SCI模型,并随机分为GM-1组(A组)和生理盐水对照组(B组)两组,每组各30只.分别于术后1、7、14、28、56 d采用Rivilin斜扳试验、改良Tarlov评分评价大鼠后肢运动功能恢复程度后处死取材,每组各时相点6只.以组织学和免疫荧光染色观察大鼠脊髓损伤的修复情况.[结果]术后7d起,Rivilin斜扳试验和Tarlov评分在A、B组间比较有显著性差异(P<0.0105),即A组功能恢复明显优于B组.术后56 d:HE染色示A组大鼠脊髓损伤处无明显空洞和瘢痕组织,有各种形态细胞形成,部分细胞有明显分化特征;B组脊髓断端被瘢痕组织填塞,可见大量炎性细胞和成纤维细胞浸润,并有较大脊髓空洞形成.免疫荧光染色发现,A组各时相点MAP-2表达呈阳性细胞数较B组高,差异有统计学意义(P <0.0105).[结论]SCI后,动物神经功能的恢复与MAP-2的表达呈一定的相关性;GM-1可通过增强脊髓损伤后MAP-2的表达来保护大鼠受损后脊髓的神经元.     

Passive biaxial mechanical properties of the rat bladder wall after spinal cord injury

PURPOSE: Changes in the mechanical properties of the bladder wall after spinal cord injury can alter bladder compliance and wall tension, leading to changes in afferent nerve activity and to abnormal reflex mechanisms. To our knowledge we report the first application of biaxial mechanical testing to the normal bladder wall and demonstrate how these properties change after spinal cord injury. MATERIALS AND METHODS: Whole bladders were harvested from mature female Sprague-Dawley rats weighing 250 to 300 gm. Test group animals underwent complete spinal cord transection at the T9 to T10 level and normal animals comprised the control group. The bladders were cut open longitudinally, the trigone and apex were removed and the remaining tissue was trimmed to a square of 9 to 13 mm. per side. Mechanical properties of the tissue sample were tested using planar biaxial testing, in which a stress was applied in the circumferential and longitudinal (base-apex) directions, and resulting axial strains were measured. RESULTS: In normal and spinal cord injured rats bladder wall tissue demonstrated isotropic mechanical behavior when equal stress levels were applied in anatomical directions. However, under nonequi-biaxial loading bladder specimens were not truly isotropic but displayed anisotropic-like behavior. Spinal cord injured tissues were consistently more compliant than normal controls. CONCLUSIONS: Biaxial mechanical testing can detect differences in normal control and hypertrophied rat bladders 10 and 14 days after spinal cord injury. These changes represent an important component of the bladder response to spinal cord injury.     

对大鼠脊髓损伤应用红花注射液的保护作用

[目的]采用重物下降打击法,制备相当于中度大鼠脊髓损伤(spinal cord injury,SCI)的模型,探讨红花注射液对SCI的保护效果。[方法]将72只SD大鼠随机分为假手术组(A组)、脊髓打击损伤组(B组)、甲基强的松龙组(methylprednisolone sodium succinate,MPSS)(C组)、红花溶液组(D组),每组18只。B、C、D组根据改良的重物撞击装置制备脊髓急性打击损伤动物模型,C、D组在脊髓打击损伤后即刻分别腹腔注射MPSS 30 mg/kg和红花溶液100 mg/kg,观察并检测4组大鼠SCI后1、24、48 h 3个不同时间点后肢运动功能评分变化以及脊髓组织病理学、组织含水量、乳酸(lactic acid,LD)含量、乳酸脱氢酶(lactic dehydrogenase,LDH)活性、超氧化物歧化酶(superoxide dismutase,SOD)活性、丙二醛(malondialdehyde,MDA)含量的改变。[结果]术后A组大鼠功能完全恢复;C组或D组与B组相比,大鼠后肢运动功能24、48 h 2个时间点均有所改善,但24 h时已出现明显差异(P<0.05)...     

Electrophysiological evidence for an antinociceptive effect of ketamine in the rat spinal cord

Background : The dissociative anesthetic ketamine also has antinociceptive effects. The mechanism and the site of action of such effect of ketamine have been, however, elusive and controversial. The present study was conducted to examine the effect of systemically administered ketamine on spinal nociceptive transmission.
Methods : We investigated and compared the effects of ketamine (0.25-8 mg/kg) on the hamstring flexor reflex in intact, lightly anesthetized rats and spinally transected rats. The opioid receptor antagonist naloxone was used to examine the involvement of opioid receptors in the actions of ketamine. Finally, the effects of ketamine on dorsal horn neuronal activity to electrical stimulation of peripheral nerves were also studied.
Results : Ketamine caused similar dose-dependent depression of the hamstring flexor reflex recorded from spinally intact rats and from spinalized rats. Even the highest dose of ketamine failed to influence the monosynaptic reflex. The depressive effect of ketamine on the flexor reflex was not reversed by naloxone. Ketamine i.v. also exerted a relatively selective inhibition of the responses of dorsal horn wide-dynamic-range neurons to C-fiber input of electrical stimulation of the plantar nerve.
Conclusions : Our present results support the notion that ketamine can exert a direct antinociceptive effect in rat spinal cord. Moreover, the data indicated that the spinal antinociceptive effect of ketamine does not involve naloxone-sensitive opioidergic mechanisms.     

The effect of intravesical electrical stimulation on bladder function and synaptic neurotransmission in the rat spinal cord after spinal cord injury

OBJECTIVE

To investigate the effects of intravesical electrical stimulation (IVES) on bladder function and synaptic neurotransmission in the lumbosacral spinal cord in the spinalized rat, as the clinical benefits of IVES in patients with increased residual urine or reduced bladder capacity have been reported but studies on the mechanism of IVES have mainly focused on bladder Aδ afferents in central nervous system‐intact rats.

MATERIALS AND METHODS

In all, 30 female Sprague‐Dawley rats were divided equally into three groups: normal control rats, sham‐stimulated spinalized rats and IVES‐treated spinalized rats. IVES was started 5 weeks after spinal cord injury (SCI) and was performed 20 min a day for 5 consecutive days. At 7 days after IVES, conscious filling cystometry was performed. Sections from the L6 and S1 spinal cord segments were examined for n ‐methyl‐d ‐aspartic acid receptor 1 (NMDAR1) subunit and γ‐aminobutyric acid (GABA) immunoactivity.

RESULTS

In IVES‐treated spinalized rats, the number and maximal pressure of nonvoiding detrusor contractions were significantly less than in sham‐stimulated spinalized rats. The mean maximal voiding pressure was also lower in IVES‐treated than in sham‐stimulated spinalized rats. IVES significantly reduced the interval between voiding contractions compared with the untreated spinalized rats. There was an overall increase in NMDAR1 immunoactivity after SCI, which was significantly lower in IVES‐treated spinalized rats. Immunoactivity of GABA after SCI was significantly lower than in the control group and was significantly higher in IVES‐treated spinalized rats.

CONCLUSION

Our results suggest that IVES might affect voiding contractions in addition to inhibiting C‐fibre activity and that IVES seems to have a more complex effect on the bladder control pathway. For synaptic neurotransmission in the spinal cord, IVES could possibly shift the balance between excitation and inhibition towards inhibition.