siRNA沉默波形蛋白表达抑制大鼠急性脊髓损伤胶质瘢痕的形成

背景:脊髓损伤后胶质瘢痕形成物理屏障抑制轴突跨越损伤部位,被认为是神经修复过程中的不利因素.目的:研究siRNA干扰波形蛋白表达对反应性星形胶质细胞的影响,探讨其在脊髓损伤后胶质瘢痕形成中的意义.方法:①选取生长良好的第3代星形胶质细胞,分为未转染组、siRNA-NC组、siRNA-Vimentin组,利用siRNA干...     

大鼠脊髓胶质瘢痕形成的规律*

背景：脊髓损伤后治疗不理想的原因是脊髓组织的囊变和胶质瘢痕的形成,因此,明确胶质瘢痕的发生发展规律具有重要意义。 目的：观察大鼠脊髓损伤后脊髓胶质瘢痕形成的空间分布、时间规律,以及轴突变化特征。 方法：采用改良Allen重物坠落法建立SD大鼠脊髓损伤模型,分别于损伤后1 d,3 d,5 d,1周,2周,4周,6周,8周,10周,12周取材。以正常饲养的大鼠作对照。 结果与结论：大鼠脊髓损伤后4周开始出现致密瘢痕增生,之后瘢痕厚度平稳下降,至损伤后10周形成光滑的囊腔壁,囊腔内无胶质纤维酸性蛋白阳性星形胶质细胞,损伤区囊腔周围的胶质瘢痕内可见密集肥大的星形胶质细胞,未见神经丝蛋白阳性轴突位于囊腔内。提示脊髓损伤后4周胶质瘢痕厚度达到高峰,囊腔与残存轴突之间开始形成机械屏障,损伤后10周瘢痕厚度趋于稳定。      

大鼠脊髓损伤后星形胶质细胞特异性表达Gc的研究

用免疫组织化学方法观察了脊髓的星形胶质细胞在损伤后出现的抗原性改变并对其改变的意义进行了探讨。实验选用Wistar大鼠 2 0只。实验组 10只 ,对脊髓 T1 0 节段进行完全横断 ;对照组 10只 ,只进行 T1 0 椎板切除术 ,不损伤脊髓。在术后第 1、3、5、7、14 d分别对 2 0只大鼠灌流固定 ,并取出 3 cm长手术段脊髓。用 anti-Galactocerebrosides( anti-Gc)和 anti-glial fibrillaryacidic protein( anti-GFAP)抗体对脊髓进行标记。结果表明 :脊髓损伤后第 7d,增生肥大的星形胶质细胞可以同时被 anti-GF AP和 anti-Gc标记 (荧光双标 )。此抗原表型改变至术后 14 d依然显现。被双标的星形胶质细胞在形态上与成熟的正常胶质细胞基本相同 ,而少突胶质细胞只为 anti-Gc单独标记。对照组脊髓星形胶质细胞和少突胶质细胞只为 anti-GFAP、anti-Gc分别标记。本实验结果提示 :大鼠脊髓受损后 ,星形胶质细胞出现 GFAP和 Gc二种抗原表型。此结果首次表明成熟哺乳动物脊髓损伤后星形胶质细胞也可出现类似少突胶质细胞特异性抗原抗体改变。这可能是星形胶质细胞对脊髓创伤的一种特异性反应。这种变化可为探索脊髓损伤区域微环境的变化对脊髓损伤修复的影响提供新的线索     

星形胶质细胞调节缺血性脑卒中的胶质瘢痕形成

背景：缺血性脑卒中是临床上主要的致死及致残性疾病之一,经血管再通获益的患者数量极其有限,故探索有效治疗手段迫在眉睫。以星形胶质细胞为主所形成的胶质瘢痕作为缺血性脑卒中的重要病理变化之一,是阻碍脑卒中后期轴突再生和神经修复的主要原因。目的：通过对缺血性脑卒中后星形胶质细胞瘢痕形成的病理过程、关键的信号调节机制和潜在治疗靶点进行分析,旨在为干预星形胶质细胞瘢痕形成以有效治疗缺血性脑卒中提供理论依据,并为促进脑卒中后康复提供新策略。方法：全面检索2010-2022年在中国知网、Pub Med和Web of Science数据库收录的相关文献,中文检索词：“缺血性脑卒中、脑缺血、星形胶质细胞、胶质瘢痕、胶质增生、星形胶质细胞增多症”,英文检索词：“Ischemic stroke,Brain ischemi*,Cerebral ischemi*,Astrocyt*,Astroglia*,Glial scar,Gliosis,Astrogliosis”,经筛选后纳入78篇文献进行综述。结果与结论：(1)星形胶质细胞在中枢神经系统稳态的维持中具有重要作用,缺血性脑卒中发生后,星形胶质细胞从静息态转变...     

人胎儿脊髓星形胶质细胞形态及分布

目的　观察星形胶质细胞在胎儿脊髓不同发育阶段形态和分布的变化。方法　在引产 19例胎儿脊髓 ,多克隆抗体GFAP ,应用SP免疫组织化学染色和图像分析。结果　星形胶质细胞在中央管、毛细血管周围密度大 ,染色深 ,环绕血管呈辐射状排列。 16W时已有少突起GFAP阳性细胞出现 ,胞体小 ,分布于脊髓白质的周边部 ,细长突起指向中央管方向 ;突起较短的细胞分布于脊髓前后正中沟两侧和中央管周围 ;灰质内阳性细胞主分布于背侧部的两侧 ,突起多而短 ,细胞核大。 2 4W时 ,多突起细胞增多 ,GFAP阳性细胞染色强度、细胞密度接近出生时水平。结论　人胎儿脊髓星形胶质细胞 16W时最多 ,2 4W时逐渐减少至出生时水平     

成年大鼠脊髓完全性横断后星形胶质细胞的时空分布及其变化

本研究以成年大鼠脊髓完全性横断模型研究反应性胶质细胞的时空分布和变化。将30只成年Wistar大鼠随机分为5组:正常对照组,T9横断伤1周、2周、4周和8周组,每组6只。利用免疫组织化学方法及图像分析系统对各组动物脊髓内星形胶质细胞的时空分布和变化进行观察和分析。结果显示:脊髓横断组胶质纤维酸性蛋白(GFAP)阳性的星形胶质细胞数目较正常对照组明显增加(P<0.05);距损伤近侧端较距损伤远侧端的GFAP阳性星形胶质细胞数目增加显著(P<0.05);脊髓横断组髓磷脂碱性蛋白(MBP)阳性的少突胶质细胞数目的时间及空间分布与正常对照组相比无统计学差异(P>0.05)。实验结果提示,星形胶质细胞是胶质瘢痕的主要成分,而少突胶质细胞在瘢痕形成过程中并非是反应活跃的成分。     

大鼠脊髓星形胶质细胞的分离与纯化

目的:纯化培养和鉴定新生大鼠的脊髓星形胶质细胞,建立稳定的星形胶质细胞培养体系,为研究创伤后慢性神经病理性痛的脊髓机制奠定基础。方法:根据成纤维细胞与星形胶质细胞生长倍增时间的差异,用阿糖胞苷(Ara-C)抑制成纤维细胞生长分裂的方法获取星形胶质细胞,用胶质原纤维酸性蛋白(GFAP)免疫荧光法对其鉴定。同时与差速贴壁培养法进行对比。结果:用Ara-C处理后,可获得较高纯度的星形胶质细胞,经鉴定其纯度可达90%以上,而对照组几乎全部为成纤维细胞。结论:Ara-C处理能彻底清除成纤维细胞,从而保持星形胶质细胞的正常生长繁殖,达到纯化星形胶质细胞的目的。     

脊髓损伤激活的星形胶质细胞在组织保护和功能恢复中的作用

神经组织损伤后相关基因的表达发生变化。在细胞分化、增生以及瘢痕形成等过程中均伴随着星形胶质细胞不同程度的改变 ( Eddleston M & Mucke L.Neuroscience,1993;5 4 :15 - 36 ;Eng L F & Ghirnikar RS.BrainPathol,1994 ;4 :2 2 9- 2 37)。星形胶质细胞能够清除细胞外液的谷氨酸和 K+,产生大量的生长因子和细胞因子并且是潜在的能量来源 ( Kettenm ann H & Ransom BR.Neuroglia,New York:Oxford U P,1995 )。虽然人们早已认识到星形胶质细胞在组织损伤后的修复过程中可能具有重要的影响 ,但到目前为止对其仍了解甚微。本实验采…     

脊髓损伤模型大鼠胶质细胞反应性增生的变化规律及意义

BACKGROUND: The nervous reconstruction and repair after spinal cord injury have become a research hotspot.     

胶质细胞移植治疗脊髓损伤的研究进展

脊髓损伤具有高发生率、高致残率、高生存率、高消耗、青壮年多发等特点。其相关治疗的研究主要是从挽救神经元的迟发型损害和死亡，促进神经元的再生和组织移植替代等方面进行。由于哺乳动物中枢神经系统损伤后期内源性修复能力有限，所以细胞移植可能是更为可行的修复途径。目前细胞移植分为神经干细胞移植、胶质细胞移植、基因修饰的细胞移植等。胶质细胞通过广泛分布的凸起构成神经组织的支架，对神经元胞体和突起有支持作用。出生后保持一定的分裂能力．受到创伤刺激时进行分裂增殖，形成胶质瘢痕。星形胶质细胞一方面位于神经细胞的附近．另一方面附着于毛细血管的终足。参与物质运输和血脑屏障的形成。此外，星形胶质细胞可能在神经元生成、突触网络形成、神经元电活动以及特异性神经系统疾病等过程中发挥着调控作用。基于上述理论．国内外进行了一系列胶质细胞移植修复脊髓损伤的实验研究．现就相关研究进展综述如下。     

脊髓损伤后的康复实践结果

背景：脊髓损伤患者的康复结果与患者的损伤程度、治疗方法、康复时间及后期治疗等多因素有关。跨学科、全面、专业的脊髓损伤康复单元能为脊髓损伤患者提供更好的恢复。目的：综合评价脊髓损伤康复单元干预或其组合干预的效果。方法：检索2003至2014年Springer及PubMed数据库,检索词：spinal cord injury,rehabilitation practice,outcomes。根据纳入排除标准,阅读标题和摘要进行初筛,选用44篇英文文献进行分析。结果与结论：许多研究采用基于实践证据的方法,识别多种康复实践方法,把信息与结果联系以评价康复干预的效果。研究显示创伤及非创伤性混合样本中的目标实现与年龄没有差异,大多数康复结果很少有性别差异。最初入院到专科脊髓损伤中心的绝大多数脊髓损伤患者的并发症通常最低,患者尽早入住到跨学科、全面、专业的脊髓损伤单位比延迟入住缩短住院总时间。脊髓损伤患者接受正规、全面的门诊医护随访,在健康感知、独立性、抑郁症没有显著差异,但特定继发情况出现频率显著减少、程度减低。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Pregabalin as a neuroprotector after spinal cord injury in rats: biochemical analysis and effect on glial cells

As one of trials on neuroprotection after spinal cord injury, we used pregabalin. After spinal cord injury (SCI) in rats using contusion model, we observed the effect of pregabalin compared to that of the control and the methylprednisolone treated rats. We observed locomotor improvement of paralyzed hindlimb and body weight changes for clinical evaluation and caspase-3, bcl-2, and p38 MAPK expressions using western blotting. On histopathological analysis, we also evaluated reactive proliferation of glial cells. We were able to observe pregabalin's effectiveness as a neuroprotector after SCI in terms of the clinical indicators and the laboratory findings. The caspase-3 and phosphorylated p38 MAPK expressions of the pregabalin group were lower than those of the control group (statistically significant with caspase-3). Bcl-2 showed no significant difference between the control group and the treated groups. On the histopathological analysis, pregabalin treatment demonstrated less proliferation of the microglia and astrocytes. With this animal study, we were able to demonstrate reproducible results of pregabalin's neuroprotection effect. Diminished production of caspase-3 and phosphorylated p38 MAPK and as well as decreased proliferation of astrocytes were seen with the administration of pregabalin. This influence on spinal cord injury might be a possible approach for achieving neuroprotection following central nervous system trauma including spinal cord injury.     

Therapeutic approaches for spinal cord injury

This study reviews the literature concerning possible therapeutic approaches for spinal cord injury. Spinal cord injury is a disabling and irreversible condition that has high economic and social costs. There are both primary and secondary mechanisms of damage to the spinal cord. The primary lesion is the mechanical injury itself. The secondary lesion results from one or more biochemical and cellular processes that are triggered by the primary lesion. The frustration of health professionals in treating a severe spinal cord injury was described in 1700 BC in an Egyptian surgical papyrus that was translated by Edwin Smith; the papyrus reported spinal fractures as a “disease that should not be treated.” Over the last two decades, several studies have been performed to obtain more effective treatments for spinal cord injury. Most of these studies approach a patient with acute spinal cord injury in one of four manners: corrective surgery or a physical, biological or pharmacological treatment method. Science is unraveling the mechanisms of cell protection and neuroregeneration, but clinically, we only provide supportive care for patients with spinal cord injuries. By combining these treatments, researchers attempt to enhance the functional recovery of patients with spinal cord injuries. Advances in the last decade have allowed us to encourage the development of experimental studies in the field of spinal cord regeneration. The combination of several therapeutic strategies should, at minimum, allow for partial functional recoveries for these patients, which could improve their quality of life.     

The role of inducible nitric oxide synthase following spinal cord injury in rat

Acute spinal cord injury (SCI) is two-step process that first involves the primary mechanical injury and then the secondary injury is induced by various biochemical reactions. Apoptosis is one of secondary SCI mechanisms and it is thought to play an important role for the delayed neuronal injury. The enhanced formation of nitric oxide (NO) via inducible nitric oxide synthase (iNOS) has been implicated in the pathogenesis of apoptosis in SCI. The level of .iNOS mRNA peaked at 6 hr after SCI and it declined until 72 hr after SCI in a rat model. Double-immunofluorescence staining revealed that iNOS positive cells were stained for ED-1, synaptophysin, GFAP, and oligodendrocyte marker. The terminal deoxynucleotidyl-transferase-mediated dUDP-biotin nick end-labeling (TUNEL) positive cell count was higher for the 72 hr post-SCI group than for the 24 hr post-SCI group. This cell count was also higher going in the caudal direction than in the rostral direction from the epicenter, and especially for the 72 hr group. Treatment with a selective iNOS inhibitor resulted in the reduction of TUNEL-positive cells at the lesion site. These findings suggest that nitric oxide generated by the iNOS of macrophages, neurons, oligodentrocytes, and astrocytes plays an important role for the acute secondary SCI that results from apoptotic cell death.     

大剂量甲基强的松龙冲击治疗成人急性脊髓损伤疗效的Meta分析

背景：一些随机对照研究试图回答大剂量甲基强的松龙冲击治疗成人急性脊髓损伤的疗效优劣问题,得出结论各不相同。 目的：大剂量甲基强的松龙冲击治疗成人急性脊髓损伤的疗效Meta分析。 方法：计算机检索PubMed、Embase、Cochrane图书馆及中国生物医学数据库、维普信息数据库、万方数据库,手工检索相关的中英文骨科杂志。收集大剂量甲基强的松龙冲击治疗成人急性脊髓损伤的对照试验,并评价纳入研究的方法学质量。统计软件用Cochrane协作网提供的RevMan 5.0。 结果与结论：共纳入9个临床对照试验。Meta分析结果表明,与传统治疗方案相比,甲基强的松龙在给药后24 h患者的神经功能恢复、肺部感染率、胃肠道反应发生率较高,而在泌尿系感染率、术后伤口不愈合率、应激性溃疡发生率方面与传统治疗差异无显著性意义。提示大剂量甲基强的松龙冲击治疗成人急性脊髓损伤时,对神经功能的恢复有较好的效果,但是有较高的肺部感染和胃肠道反应的发生,所以在今后的治疗过程中要尽可能的避免肺部感染和胃肠道反应的发生。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Abdominal ultrasonography findings in patients with spinal cord injury in Korea

This is a retrospective study of 500 patients with spinal cord injury who underwent abdominal ultrasonography as a routine screening test from 2000 to 2003. We analyzed the results according to the different abdominal organ systems. Among the 500 cases, 226 (45.2%) showed abnormal findings. 98 cases of abnormal findings in the liver included 75 of fatty liver and 13 of mass. The 88 cases of abnormal findings in the bladder included 56 of bladder wall thickening, 14 of cystitis and 10 of urinary stone. The 35 cases of abnormal findings in the kidney included 19 of renal cyst and 6 of pelvic dilatation. The 35 cases with gallbladder abnormalities included 19 with gallstones and 11 with biliary sludge. Excluding the cases with bladder wall thickening, there were still 170 cases with abnormal ultrasonographic findings. Abdominal sonography seems to be a useful tool in detecting hidden intraabdominal pathologies in patients with spinal cord injury.     

舒芬太尼预处理对脊髓损伤模型小鼠的神经保护

BACKGROUND: Sufentanil exerts protective effects on tissues, but its roles in the repair of nervous system injury and the underlying mechanism are still unknown.     

不同时期局部移植人脐带间充质干细胞修复脊髓损伤的组织学观察

BACKGROUND:Spinal cord injury (SCI) is a disease causing a variety of motor and sensory dysfunctions, abnormal muscle tone and pathological reflex. Clinically, human umbilical cord mesenchymal stem cell transplantation has become an employed therapy for SCI. OBJECTIVE:To investigate the effect of local transplantation of human umbilical cord mesenchymal stem cells in different time after spinal cord injury in rats. METHODS:100 SPF male adult Sprague-Dawley rats were randomly divided into five groups: blank control group, SCI group, post-SCI 3-, 7-, 21-day transplantation groups (n=20 per group). Animal models of T₁₀ SCI were made by Allen’s method in the latter four groups, and rats in the three transplantation groups were given HUCMSCs transplantation at 3, 7, 21 days after SCI, respectively. RESULTS AND CONCLUSION:Compared with the SCI group, improved motor function scores, decreased interleukin-2 level, and increased serum interleukin-10 level were observed in the three transplantation groups at 49 days after modeling, indicating SCI was improved significantly in the three transplantation groups, especially in the post-SCI 7-day transplantation group. These findings suggest that human umbilical cord mesenchymal stem cell transplantation for SCI repair improves the movement function of rats, and cell transplantation at 7 days after modeling has achieved best outcomes.     

他莫昔芬对急性脊髓损伤模型大鼠的神经保护

BACKGROUND:Tamoxifen has been found to exert neuroprotection by reducing cerebral hemorrhage and edema surrounding the injured site of the spinal cord. OBJECTIVE:To investigate the neuroprotective effect of tamoxifen on rat acute spinal cord injury and the underlying mechanism. METHODS: Sixty Sprague-Dawley rats were equivalently randomized into five groups, and modeled into spinal cord injury at T10 level using modified Allen’s weight-drop method (70 g/cm), except those in sham operation group. At 30 minutes after modeling, all rats were given the intraperitoneal injection of 2.5, 5.0 and 10 mg/kg tamoxifen or same amount of normal saline, once daily. Basso, Beattie, Bresnahan (BBB) scores were recorded at 24, 48 and 72 hours after surgery. The injured spinal cord was removed at 72 hours to observe its edema. Meanwhile, the levels of interleukin-1β, interleukin-10 and tumor necrosis factor-α, as well as Caspase-3 activity were detected by ELISA; the protein levels of nuclear factor-κB p65, phosphorylated I-κBα and Caspase-3 were detected by western blot assay. RESULTS AND CONCLUSION: Compared with the model group, the hind limb function in the tamoxifen groups was significantly improved. Tamoxifen significantly decreased the water content in the rat spinal cord and inhibited spinal cord edema at 72 hours after surgery. ELISA results showed that tamoxifen significantly reduced the activity of interleukin-1β, interleukin-10, tumor necrosis factor-α and Caspase-3 (P < 0.05). Western blot assay revealed that tamoxifen significantly downregulated the expression levels of nuclear factor-κB p65, phosphorylated I-κBα and Caspase-3. These results suggest that tamoxifen protects against spinal cord injury via suppressing inflammatory response and apoptosis-associated proteins. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程