大鼠脊髓T3节段全横断后损伤节段神经元超微结构的观察

目的：观察大鼠脊髓T3节段全横断后损伤节段神经元超微结构的变化。方法：48只大鼠随机分为假手术组和脊髓T3完全横断损伤1d组、2d组、7d组、14d组、21d组,每组8只。各组大鼠BBB运动功能评分后取损伤节段脊髓,透射电镜观察脊髓神经元超微结构的变化。结果：脊髓损伤（SCI）各组大鼠的BBB评分明显低于假手术组（P ＜0.01）;随着损伤时间的延长,SCI各组大鼠的BBB评分逐渐升高（P ＜0.01）。电镜下观察,大鼠脊髓神经元在损伤后2d病理变化最为严重,表现为神经元明显肿胀,线粒体等细胞器严重受损,核固缩,染色质凝集;损伤后7d时,脊髓神经元的病理变化有所减轻;损伤后21d,脊髓神经元的病理变化明显减轻,甚至接近正常。结论：大鼠脊髓损伤后2d脊髓神经元的病理变化最为严重,后逐渐减轻。     

嗅鞘细胞移植联合应用GDNF对大鼠脊髓损伤的治疗作用

目的：研究嗅鞘细胞（OECs)移植并联合应用胶质细胞源性神经营养因了（GDNF）对脊髓损伤的保护和促进再生作用。方法；建立成年SD大鼠脊髓T8半横断损伤模型，将体外培养纯化的OECs植入脊髓损伤处，同时局部应用GDNF，采用斜板试验和BBB联合功能评分观察大鼠运动功能恢复情况，并通过辣根过氧化物酶逆行示踪技术评价OECs和GDNF对神经元存活和纤维再生的影响。结果：（1）OECs和GDNF联合应用时对皮质和红核神经元有逆行性保护作用；可促进脊髓下行传导束再生，肢体运动功能恢复，（2）OECs和GDNF联合应用组对脊髓损伤的修复作用最强，高于GDNF或OECs单用组。结论：联合应用GDNF和OECs在脊髓损伤修复治疗中具有协同作用。     

神经干细胞移植治疗脊髓损伤的实验研究

目的：观察神经干细胞（NSCs）移植对脊髓损伤（SCI）大鼠功能恢复的作用。方法：30只Wistar大鼠随机分为对照组、损伤组和移植组,每组10只;损伤组和移植组制作成L4平面的脊髓全横断模型,将培养的大鼠NSCs悬液注入移植组损伤脊髓处,损伤组注射等量的生理盐水。术后2mo,采用BBB评分、皮层体感诱发电位（CSEP）和辣根过氧化物酶（HRP）逆行示踪技术观察大鼠脊髓运动和传导功能的恢复程度。结果：术后2mo,BBB评分损伤组、移植组大鼠有所恢复,但都未达到正常水平,其中移植组的大鼠恢复较好,评分较高,与损伤组有统计学差异。SCI后,损伤组、移植组的CSEP波消失,术后2mo移植组的波形有所恢复,但潜伏期延长。对照组脊髓前角可见到许多HRP标记阳性神经元,损伤组未见阳性神经元,移植组可见有阳性神经元,但数目较对照组少。结论：NSCs脊髓内移植能促进损伤脊髓运动和传导功能的部分恢复。     

甲基强的松龙对大鼠脊髓损伤后微管相关蛋白-2表达的影响

目的研究甲基强的松龙对大鼠脊髓损伤后微管相关蛋白-2（MAP-2）表达的影响。方法健康成年wistar大鼠66只,随机取6只作为正常对照组,60只制作成脊髓打击伤动物模型。模型大鼠随机分为甲基强的松龙组和治疗对照组,每组30只,分别于伤后1、3、7、14和28d取材,应用免疫组织化学方法观察MAP-2的表达,用改良的Tarlov评分观察大鼠脊髓损伤后运动功能的恢复情况。结果大鼠脊髓损伤后7和14d,甲基强的松龙组MAP-2的表达强于治疗对照组;大鼠脊髓损伤后7、14和28d,甲基强的松龙组Tarlov评分大于治疗对照组。结论甲基强的松龙能促进大鼠损伤脊髓表达MAP-2,并能促进大鼠脊髓损伤后运动功能的恢复。     

大鼠Allen’s脊髓损伤模型的建立及评价

目的：建立并评价大鼠Allen’s脊髓损伤模型,为进一步研究脊髓损伤机制提供实验动物参考。方法：SD大鼠60只,体重230-270 g,随机分为空白组（20只）,手术组（20只）,功能评价组（20只）。空白组不做任何处理,手术组和功能评价组都采用Allen’s法致伤。功能评价组作手术前后的BBB功能评分和斜板试验,空白组与手术组作病理切片HE染色。结果：功能评价组术前斜板度数和BBB评分均高于术后（P〈0.05）。手术组术后1 d脊髓神经元发生退变和坏死。结论：本实验建立的大鼠Allen’s脊髓损伤模型方法可靠,易于复制,可用于脊髓损伤的相关研究。     

防己黄芪汤加味治疗大鼠脊髓半横断损伤

目的：研究中药复方对急性脊髓半切损伤的治疗疗效及对脊髓组织病理改变及星型胶质细胞的影响,并探讨其作用机制。方法：65只大鼠随机取5只作为空白组,余60只大鼠脊髓损伤造模后随机分为损伤、中药和甲基强的松龙三组,各20只。损伤组造模后不用药,中药组造模后予中药治疗,甲基强的松龙组造模后予甲基强的松龙治疗。于术后24小时、1、2、3、4周观察三组大鼠脊髓运动功能恢复、光密度测定及脊髓组织学改变。结果：中药组及甲基强的松龙组脊髓运动功能及光密度评分与损伤组相比有统计学意义（P〈0.01）;中药组病变与损伤组相比,病损缩小、空泡面积减少。中药组与甲基强的松龙组后期脊髓运动功能评分无统计学差异（P〉0.05）。结论：中药组在改善脊髓微环境、保护神经组织免受损害、促进间质细胞生长上发挥其有益作用,抑制胶质细胞的过度增生,从功能和形态学上证实了中药治疗的有效性。     

CNTF对大鼠脊髓损伤后逆行性红核神经元损伤的保护作用

目的：研究睫状神经营养因子（CNTF）对脊髓损伤后逆行性红核神经元损伤的保护作用和促进肢体功能恢复的作用。方法：将大鼠分为假手术组，生理盐水（NS）组和CNTF组，用Allen改良法撞击致SD大鼠脊髓T10损伤，经蛛网膜下腔留置导管注射NS和CNTF；术后每周进行一次BBB评分和斜板试验的行为学评分；6周时采用辣根过氧化物酶（HRP）逆行示踪技术标记红核神经元，取红部位脑组织作冰冻切片，应用四甲基联苯胺（TMB）呈色反应显示脊髓损伤后红核神经元存活率，并进行图像数据的分析。结果：（1）CNTF组行为功能高于NS组；（2）CNTF组HRP标记红核神经元数目多于对照组。结论：CNTF可减轻脊髓损伤后的逆行性损伤，而对上行运行神经元发挥保护作用，从而对肢体功能的恢复起到促进作用。     

电针治疗对大鼠脊髓半离断损伤的影响

目的：研究电针刺激对大鼠脊髓半离断损伤的治疗效果,探究脊髓损伤后督脉（“大椎、命门”）电针治疗对大鼠脊髓半离断损伤右肢运动功能的影响。方法对20只SD大鼠进行T7～T9脊髓半离断损伤,术后,随机分为对照组（ A组）、督脉电针组（B组）,每组10只,A组进行饲料单纯喂养,B组则同时予以电针刺激治疗。通过BBB运动功能评分法、组织形态检测等方法来观察电针刺激督脉对脊髓半离断损伤的作用,其分别于第1、3、7、14、21天采用BBB运动功能评分法评价并对比两组大鼠右肢运动功能恢复水平;21天后,处死大鼠取其损伤段1 cm左右脊髓制片,采用组织形态检测来观察其组织病理变化。结果 A、B两组大鼠术后第1、3、7、14、21天BBB运动功能评分逐渐增高,B组BBB评分明显高于A组（ P＜0．05）。HE染色可见B组较A组脊髓组织结构清晰,有较多神经元存活,其无明显水肿、坏死,无出血、炎性细胞浸润和瘢痕组织形成,可见大量正常神经元。结论电针刺激督脉治疗能促进大鼠脊髓半离断损伤右肢运动功能的部分恢复。     

金腰带联合甲基强的松龙对脊髓损伤大鼠行为学及脑源性神经营养因子表达的影响

目的探讨金腰带和甲基强的松龙联合应用对脊髓损伤（SCI）大鼠的疗效及其对脑源性神经营养因子（BDNF）表达的影
响,为临床治疗脊髓损伤提供理论依据。方法将雌性SD大鼠30只随机分为：（1）假手术组;（2）脊髓挫伤组;（3）甲基强的松龙
治疗组（术后8 h内肌注50 mg/kg,此后每天甲基强的松龙肌注量减少10 mg/kg）;（4）金腰带治疗组（50 mg/kg,每日1次灌胃）;
（5）金腰带和甲基强的松龙联合治疗组（两种药物及给药方法相加）。采用BBB评分系统对后肢运动功能进行评分。对大鼠脊髓损伤
术后4周的脊髓切片进行BDNF免疫组化染色,观察阳性细胞定位、分布以及数量变化。用方差分析进行多个样本间比较。结
果BDNF免疫反应阳性产物主要分布于脊髓灰质腹角和背角神经元。联合治疗组BDNF阳性神经元数量以及BBB评分分值
较假手术组、脊髓挫伤组、甲基强的松龙治疗组和金腰带治疗组均显著增高, 差异有统计学意义（P<0.05）。结论甲基强的松龙
和金腰带联合治疗脊髓损伤的疗效优于单纯甲基强的松龙治疗组和金腰带治疗组,其发挥疗效可能与上调BDNF表达有关。
     

预防使用米诺环素对胸段脊髓全横断大鼠运动功能恢复的影响

目的：观察预防使用米诺环素对脊髓全横断大鼠运动功能的作用。方法：30只雄性Wistar大鼠随机分为3组,假手术组、脊髓横断模型组和米诺环素预处理组,每组10只大鼠。米诺环素预处理组大鼠术前腹腔注射米诺环素（90mg/kg）。各组大鼠于术后1、7、14、21d应用改良Tarlov评分和BBB评分方法对大鼠后肢行为功能进行评价。第22d将所有实验大鼠处死后取损伤段脊髓,行HE染色。结果：术后第1d,脊髓横断模型组和米诺环素预处理组大鼠Tarlov评分障碍率、BBS评分比较无明显差别（P〉0.05）;米诺环素预处理组术后第7d到第21dTarlov评分障碍率明显低于脊髓横断模型组,BBB评分明显高于脊髓横断模型组（P〈0.01）。HE染色结果显示,脊髓横断模型组大鼠脊髓出现明显病理变化,米诺环素预处理组大鼠脊髓的病理变化明显减轻。结论：预防使用米诺环素可明显促进胸段脊髓横断大鼠运动功能的恢复。     

X线照射促进大鼠脊髓损伤区结构及损伤后功能的恢复

目的探讨X线照射能否促进大鼠压迫型脊髓损伤区结构及损伤后功能的恢复。方法70只Sprague-Dawley大鼠均接受胸2节段的脊髓损伤,然后被随机分为2组,照射组在损伤后14d接受X线照射,对照组则不接受任何治疗。所有大鼠分别在脊髓损伤后第3、7、14、21、28、35和42天接受后肢运动、斜板及后肢疼痛反射等3项脊髓功能测定,43d将所有实验大鼠处死后取出脊髓损伤标本,进行组织学检查。结果70只实验鼠中符合设计要求的共62只,其中照射组32只,对照组30只。X线照射组大鼠的后肢运动恢复及斜板测定均明显优于对照组,两者之间的差异有显著性(P<0.01),而疼痛反射两组间差异无显著性。照射组大鼠脊髓损伤部位的水肿及坏死区域明显少于对照组,而神经元数目则明显多于对照组。结论X线照射可通过改善脊髓损伤区结构来促进大鼠脊髓损伤后运动功能的恢复。     

FUNCTIONAL AND STRUCTURAL RECOVERY OF INJURED SPINAL CORD FOLLOWING DELAYED X-IRRADIATION IN RATS

PRIOR studies have shown structuraland functional recovery following spinalcord injury (SCI)variousmodels.1Specifically, structural recoverafterX-irradiation forunilaterally transected adultrat spincord has been demonstrated.2This study was designed tdete…     

条件培养基局部移植对大鼠脊髓损伤的修复作用

目的：观察骨髓间充质干细胞的条件培养基(BMSC-CM)局部移植对脊髓损伤修复的作用。方法采用改良的Allen 装置制作大鼠 T9脊髓损伤模型,随机分为对照组(脊髓损伤后局部微量注射生理盐水10μl)和实验组(脊髓损伤后局部注射 BMSC-CM 10μl),两组均采用 BBB 评分、斜板实验法观察大鼠运动功能恢复情况,苏木精-伊红(HE)染色观察脊髓损伤处血管生长及空洞体积的改变情况。结果实验组大鼠的 BBB 后肢功能评分、斜板倾斜角度优于对照组(P <0．05),脊髓损伤处空洞体积明显小于对照组,血管直径大于对照组。结论 BMSC-CM 能够减小脊髓损伤处的空洞体积和促进血管生长,从而促进脊髓损伤后神经功能的恢复。     

Human neural stem cells promote corticospinal axons regeneration and synapse reformation in injured spinal cord of rats

Background Axonal regeneration in lesioned mammalian central nervous system is abortive, and this causes permanent disabilities in individuals with spinal cord injuries. This paper studied the action of neural stem cell （NSC） in promoting corticospinal axons regeneration and synapse reformation in rats with injured spinal cord. Methods NSCs were isolated from the cortical tissue of spontaneous aborted human fetuses in accordance with the ethical request. The cells were discarded from the NSC culture to acquire NSC-conditioned medium. Sixty adult Wistar rats were randomly divided into four groups （n=15 in each）： NSC graft, NSC medium, graft control and medium control groups. Microsurgical transection of the spinal cord was performed in all the rats at the T11. The NSC graft group received stereotaxic injections of NSCs suspension into both the spinal cord stumps immediately after transection; graft control group received DMEM injection. In NSC medium group, NSC-conditioned medium was administered into the spinal cord every week; NSC culture medium was administered to the medium control group. Hindlimb motor function was assessed using the BBB Locomotor Rating Scale. Regeneration of biotin dextran amine （BDA） labeled corticospinal tract was assessed. Differentiation of NSCs and the expression of synaptophysin at the distal end of the injured spinal cord were observed under a confocal microscope. Group comparisons of behavioral data were analyzed with ANOVA. Results NSCs transplantation resulted in extensive growth of corticospinal axons and locomotor recovery in adult rats after complete spinal cord transection, the mean BBB scores reached 12.5 in NSC graft group and 2.5 in graft control group （P〈 0.05）. There was also significant difference in BBB score between the NSC medium （11.7） and medium control groups （3.7, P〈 0.05）. BDA traces regenerated fibers sprouted across the lesion site and entered the caudal part of the spinal cord. Synaptophysin expression colocalized with BDA positive axons and neurons distal to the injury site. Transplanted cells were found to migrate into the lesion, but not scatter along the route of axon grows. The cells differentiated into astrocytes or oligodendrocytes, but not into the neurons after transplantation. Furthermore, NSC medium administration did not limit the degree of axon sprouting and functional recovery of the injured rats compared to the NSC graft group. Conclusions Human embryonic neural stem cells can promote functional corticospinal axons regeneration and synapse reformation in the injured spinal cord of rats. The action is mainly through the nutritional effect of the stem cells on the spinal cord.     

Effects of Co-grafts Mesenchymal Stem Cells and Nerve Growth Factor Suspension in the Repair of Spinal Cord Injury

Endogenous repair ability of mammalsisli mit-ed after the damage of central nervous system.It isdifficult to produce the new neurons for the dam-aged adult central nervous system,and neither canthe damaged systemstart the functional axonal re-generation. At present ,a good many methods arebelieve to be able toi mprove the damagedlocal mi-croenvironment , such as cell transplantation andprovision of exogenous neurotrophic factors ,whichare ai med at promoting the regeneration of thedamaged spin…     

超早期高压氧治疗对脊髓全横断大鼠脊髓水肿及后肢运动功能的影响

目的 探讨超早期高压氧治疗对脊髓全横断损伤大鼠脊髓水肿及后肢运动功能的影响.方法 (1)55只3月龄健康雌性SD大鼠随机分为假手术组(Sham组,15只)、模型组(CSCT组,20只)及高压氧组(HBO组,20只).Sham组仅行椎板切除术,其余2组均行T10椎板水平脊髓完全横向切断术.(2)HBO组于术后第3小时置于动物舱内开始高压氧治疗,10天1疗程,共3疗程,其中第1疗程每天2次,第2、3疗程均每天1次,每疗程休息6 d.(3)分别于建模型后第1～6周末.采用BBB运动功能评分法评价并比较两组大鼠后肢运动功能恢复程度;术后6周末取材,观察各组受损脊髓节段组织含水量变化.结果 CSCT组及HBO组大鼠术后第1-6周末BBB运动功能评分逐渐增高,HBO组在2～6周末的BBB运动功能评分均明显高于CSCT组(P<0.05);术后6周末,CSCT组受损脊髓节段组织含水量较Sham组明显增加(P<0.01),HBO组受损脊髓节段组织含水蛩较CSCT组有所减少(P<0.05).结论 超早期高压氧治疗可可减轻脊髓全横断损伤大鼠脊髓组织水肿,促进后肢运动功能的部分恢复,对脊髓有保护作用.

Abstract：

Objective To evaluate the effect of ultra-early hyperbaric oxygenation on spinal edema and hind limb motor function in rats with complete spinal cord transection. Methods Fifty-five healthy 3-month-old female SD rats were randomly divided into sham-operated group (n=15), complete spinal cord transection group (CSCT group, n=20) and hyperbaric oxygen group (HBO group, n=20). The rats in the sham-operated group underwent only laminectomy, while those in the other 2 groups underwent CSCT at the T10 level. The rats in HBO group were placed in an oxygen chamber 3 h after the operation for 10 days as a treatment course, and 3 treatment courses were administered at the interval of 6 days. In the first treatment course, 2 hyperbaric oxygenation sessions were given daily, and in the following 2 course, only 1 session was given daily. The recovery of hindlimb motor function was evaluated using the open-field Basso-Beattie-Bresnahan (BBB) scoring system once a week for 6 weeks. All the rats were sacrificed 6 weeks after the operation to measure the water content in the injured tissues. Results The BBB scores of CSCT group and HBO group gradually increased with the passage of time after the operation, and from week 2 to week 6, HBO group had significantly higher scores than CSCT group (P<0.05). The water content was markedly increased in CSCT group at 6 weeks after the operation as compared with that in the sham-operated group (P<0.01), and significantly reduced in HBO group in comparison with that in the CSCT group (P<0.05). Conclusion Ultra-early HBO can suppress spinal cord edema and promote hindlimb locomotor recovery in rats with complete spinal cord transection.     

针刺对大鼠脊髓损伤后脊髓前角运动神经元功能的影响

目的探讨针刺对脊髓损伤(SCI)大鼠前角运动神经元功能恢复的影响。方法成年雄性SD大鼠60只,改良Allen法压迫致伤T8、T9,随机分为实验组和对照组,伤后分别给予电针刺激大鼠双下肢足三里和悬钟、伏兔和三阴交穴,取治疗2,4,6周伤段脊髓进行PCR扩增和组织切片。利用RT-PCR技术检测伤区脊髓胶质源性神经营养因子(GDNF)mRNA基因体内表达的变化;应用尼氏染色、酶组织化学染色方法观察大鼠SCI后伤区脊髓前角运动神经元存活的数目和乙酰胆碱酯酶(AChE)的变化;采用斜板试验和BBB评分法观察大鼠后肢运动功能恢复情况。结果实验组较对照组AChE活性显著增加(P<0.01);实验组脊髓前角大、中型神经元的数目较对照组明显增加(P<0.05);实验组GDNF mRNA基因体内表达较对照组明显增加(P<0.01);大鼠SCI 3周后针刺治疗组后肢运动功能评分均明显高于对照组(P<0.05)。结论针刺治疗对脊髓损伤后前角运动神经元的功能恢复有一定的促进作用。     

Changes in the expression of platelet-derived growth factor in astrocytes in diabetic rats with spinal cord injury

Background Prospective mortality studies in the United States revealed that the mortality was elevated in diabetics compared to normal individuals following chronic spinal cord injury （SCI）. Our study was conducted to investigate the levels of platelet-derived growth factor （PDGF） of astrocytes in SCI in streptozotocin （STZ）-induced diabetic rats. Methods Thirty male Sprague-Dawley （SD） rats were randomly divided into 3 groups： SCI group, diabetic SCI group, and sham operation control group. We employed STZ-induced diabetic SD rats and a weight-drop contusion SCI model. The rats were sacrificed on day 7 after the induction of SCI. Immunohistochemistry and Western blotting analysis were used to detect the PDGF expression level. Basso, Beattie and Bresnahan locomotor rating scale （BBB） was also used to evaluate the neurological recovery level of the rats. Results PDGF positive astrocyte numbers were significantly higher and PDGF staining was more intensive in astrocytes in the SCI group than in the diabetic SCI group （P〈0.05）. The diabetic SCI group showed a slower recovery of motor function with a lower BBB score 7 days after acute spinal injury. Conclusions PDGF is an important factor for the recovery of neurological function after acute spinal injury and hyperglycemia in diabetic rats could depress the expression of PDGF in injured spinal cord. This may help to explain the slower recovery and higher mortality in diabetics after SCI.