干细胞的研究进展

在美国《Ｓｃｉｅｎｃｅ》杂志评选出的１９９９年度１０大科学进展中，干细胞（ｓｔｅｍ ｃｅｌｌ， ＳＣ）的研究工作格外令人瞩目。这不仅因为在过去短短的一年里，科学家们在干细胞的功能研究方面发表了１０余篇具有里程碑意义的论文，更重要的是，这些突破性进展使得生物学家和医学家们更进一步地认识到：ＳＣ作为一类既有自我更新能力、又有多分化潜能的细胞，具有非常重要的理论研究意义和临床应用价值［１］。这些成果一方面揭示了许多有关细胞生长和发育的基础理论难题；另一方面，可望将其用于创伤修复、神经再生和抗衰老等临床医…     

干细胞移植治疗肝脏疾病的研究进展

背景：干细胞移植治疗肝病是近年来众多学者研究的热点,干细胞移植的基本理论和临床应用研究都取得了很大的进展。 目的：对干细胞移植的理论依据、干细胞来源、移植方式、实验与临床研究、存在问题及前景进行简要综述。 方法：应用计算机检索中国学术期刊全文数据库(CNKI)和Pubmed数据库中2001-01/2011-11关于干细胞移植治疗肝病的文章,检索主题词“干细胞,移植,肝脏疾病,肝损伤”或“stem cell,transplantation,hepatic disease, hepatic injury”。初检索到192篇文献,据纳入标准保留31篇进行分析、综述。 结果与结论：干细胞来源充足,容易获取,可以体外增殖培养,干细胞移植操作简单,安全性高,尤其自体干细胞移植可完全避免移植排斥反应。但自体干细胞移植肝脏疾病的安全性和有效性尚无公识,需更长期的观察。     

胚胎神经干细胞移植治疗阿尔茨海默病

BACKGROUND:More recently, stem cell therapy has become an issue of concern. Exogenous neural stem cell transplantation brings new hope for the treatment of nervous system injury by self-replication and differentiation to complement and replace damaged or dead nerve cells. OBJECTIVE:To explore the therapeutic efficacy of neural stem cell transplantation on Alzheimer’s disease. METHODS:Thirty APP/PS1 mice with Alzheimer’s disease were randomly assigned into model group, cell solution transplantation group or cell transplantation group (n=10 per group). Another 10 C57BL/6 mice were selected as controls. Embryos of C57BL/6 mice at 18 embryonic days were taken to make neural stem cell suspension followed by transfection using lentiviral vectors carrying GFP gene at different multiplicities of infection (1, 5, 10, 15, 20). Afterwards, GFP-transfected neural stem cells were implanted into the hippocampus of Alzheimer’s disease mice in the cell transplantation group, while the same volume of complete medium was injected into the hippocampus of mice in the cell solution transplantation group. Morris water maze test was performed at 2 weeks after cell transplantation, and brain tissues of mice was taken and detected histologically at 4 weeks after cell transplantation. RESULTS AND CONCLUSION:Compared with the control group, the escape latency was significantly higher, and the number of crossings over the target quadrant was lower in the other three groups (P < 0.05). Compared with the cell solution transplantation and model groups, in contrast, the escape latency was significantly lower, and the number of crossings over the target quadrant was significantly higher in the cell transplantation group (P < 0.05). Four weeks after transplantation, more intact neurons were found in the cell transplantation group as compared with the model group. These findings indicate that neural stem cell transplantation can improve behavior and morphology performance of mice with Alzheimer’s disease.     

干细胞移植治疗系统性硬化的研究进展

系统性硬化(systemic sclerosis,SSc)的治疗一直是临床上的难题,现有的治疗手段难以满足患者需求。造血干细胞移植(hematopoietic stem cell transplantation,HSCT)在临床上的发展已相对成熟,近年来其在SSc上的临床试验取得了突破进展,但移植相关并发症问题一直有待...     

神经干细胞移植治疗老年痴呆

BACKGROUND:Neural stem cell transplantation has been used to treat a series of brain injury diseases, such as cerebral palsy, but its effect on Alzheimer’s disease is rarely reported. OBJECTIVE:To observe the effect of neural stem cell transplantation on the behavior and immune regulating system of Alzheimer’s disease rats.  METHODS:Thirty-five Sprague-Dawley rats were enrolled to make a postcerebral incision and given hippocampal injection of amanita phalloides acid to establish rat models of Alzheimer’s disease. Another 10 rats were only given hippocampal injection of normal saline after preparation of postcerebral skin incision as sham operation group. Then 32 successful rat models were randomly divided into two groups (n=16 per group): rats in experimental group were administrated hippocamal injection of 5×109/L allogeneic neural stem cell suspension; those in model group were given no injection. Five-day Morris water maze test was conducted at 4 weeks after transplantation. At 1 week after Morris water maze test, levels of interleukin-1 and interleukin-10 in the cerebral homogenate were detected, as well as pathological changes of brain tissues were observed in the three groups. RESULTS AND CONCLUSION:Compared with the model group, the abilities of cognition and memory were significantly higher in the sham operation group (P < 0.01), and the abilities of spatial learning and memory were significantly higher in the experimental group (P < 0.05, P < 0.01). Levels of interleukin-1 and interleukin-10 in the model group were significantly higher than those in the sham operation group (P < 0.01) but significantly lower than those in the experimental group (P < 0.01). Besides, the number of neurons in the model group was obviously less than that in the experimental and sham operation group. These results indicate that neural stem cell transplantation supplements and protects neurons against Alzheimer's disease in rats, thereby significantly improving the learning and memory ability.     

同基因与异基因胚胎肝干细胞移植治疗小鼠肝硬化

背景：胚胎肝干细胞移植免疫相关研究较少,同基因与异基因胚胎肝干细胞移植对小鼠肝硬化的治疗作用,目前尚不清楚。 目的：观察同种同基因与同种异基因胚胎肝干细胞移植对小鼠肝硬化的治疗作用,以及治疗过程中免疫排斥反应发生情况。 方法：采用Ⅳ型胶原酶消化法分离纯化BALB/c与C57BL/6胚胎肝干细胞。104只健康BALB/c小鼠随机分为4 组：正常对照组不予任何处理;肝硬化组、同种同基因移植组、同种异基因移植组腹腔注射四氯化碳石蜡油溶液复制肝硬化模型,16周后分别经其尾静脉注射生理盐水,等量同种同基因胚胎肝干细胞和同种异基因胚胎肝干细胞。在移植4周后比较各组受体小鼠存活情况、肝功能恢复情况、肝纤维化程度、免疫细胞(CD4⁺T、CD8⁺T、NK、NKT)数目及比值、肝脏病理学变化。 结果与结论：同种同基因移植组和同种异基因移植组生存率均为100%,与肝硬化组小鼠存活率67%相比差异有显著性意义(P < 0.05);各组肝功能和肝纤维化指标差异无显著性意义(P > 0.05)。各组免疫学指标比较差异无显著性意义(P > 0.05)。肝脏组织病理学显示肝组织修复：同种异基因移植组>同种同基因移植组>肝硬化组。因此,经尾静脉移植胚胎肝干细胞能提高肝硬化小鼠的生存率、减轻肝细胞坏死程度;同种同基因与同种异基因胚胎肝干细胞移植未发现免疫排斥,对小鼠肝硬化有一定的治疗作用。     

自体骨髓干细胞移植治疗肝硬化的疗效观察

治疗终末期肝病,原位肝移植是最理想的手段,然而供体短缺、手术损伤大、术后的免疫排斥反应以及费用高昂等问题限制了肝移植技术的发展和临床应用。自体骨髓干细胞移植操作简便、有效、侵入性小且并发症少,具有重要的临床意义。我科2009年9月至2011年1月共收治严重肝硬化失代偿期患者8例,经自体骨髓干细胞移植,治疗效果良好     

干细胞移植治疗心肌坏死的基础与临床研究

干细胞治疗心肌坏死是一种新的很有前景的治疗手段,但其机制尚不十分清楚。我国目前临床研究发现干细胞移植可以明显改善急性心肌梗死及梗死后心衰心脏功能,但由于关于干细胞移植的安全性还没有定论,因此还需慎重对待。     

糖原合成酶激酶3β过表达心肌干细胞移植治疗心肌梗死

BACKGROUND:The mechanism and effect of glycogen synthase kinase 3β (GSK-3β) in the differentiation of cardiac stem cells into cardiomyocytes are still unclear, although GSK-3β is closely related to the life activities of cells. OBJECTIVE:To investigate the changes of GSK-3β expression in the treatment of myocardial infarction in rats undergoing cardiac stem cell transplantation. METHODS:The isolation and culture of cardiac stem cells were performed in 10 neonatal rats. Lentivirus overexpressing GSK-3β or LacZ (control) was constructed and transferred into cardiac stem cells. Animal model of myocardial infarction was made in 30 Sprague-Dawley rats. Six weeks after model preparation, rat models were assigned into GSK-3β, LacZ or PBS group. GSK-3β or LacZ overexpressing cardiac stem cell solution or PBS in equal volume was injected into the rat myocardium, respectively. Four weeks after transplantation, the cardiac function and myocardial collagen production in rats were detected and compared. RESULTS AND CONCLUSION:Compared with the other two groups, the left ventricular ejection fraction was significantly higher, and the left ventricular end diastolic diameter was significantly lower in the GSK-3β group (P < 0.05). Hydroxyproline content, type I collagen mRNA, and type III collagen mRNA expression were significantly lower in the GSK-3β group than the other two groups (P < 0.05). Findings from Masson staining showed that the content of blue-stained collagen was significantly lower in the GSK-3β group than the LacZ group. Moreover, lowest myocardial infarction size was found in the GSK-3β group (P < 0.05). All these experimental findings show that GSK-3 overexpression plays a positive role in promoting the therapeutic effect of cardiac stem cell transplantation.     

静脉移植骨髓间充质干细胞对心肌病心力衰竭心功能的影响

BACKGROUND:Bone marrow mesenchymal stem cell transplantation can effectively improve decreased cardiac function caused by heart failure, but there is a lack of research about the effect in bone marrow mesenchymal stem cell transplantation on cardiac function in heart failure induced by cardiomyopathies. OBJECTIVE:To explore the effect of bone marrow mesenchymal stem cell transplantation on cardiac function in patients with cardiomyopathies accompanied by heart failure. METHODS:Totally 40 Sprague-Dawley rats were enrolled, and bone marrow mesenchymal stem cells were isolated from 10 rats, and the remaining rats were equivalently randomized into normal, model and stem cell transplantation groups. Then rats in the model and stem cell transplantation groups were given intraperitoneal injection of hydrochloric acid doxorubicin to prepare cardiomyopathy-induced heart failure models. At 7 days after modeling, the stem cell transplantation group was treated with bone marrow mesenchymal stem cells through intravenous transplantation, and the model group was treated with equal amount of DMEM medium. Four weeks later, cardiac function of each rat was detected, and the cell survival and differentiation were observed by immunofluorescence method. RESULTS AND CONCLUSION:At 4 weeks after transplantation, compared with normal and stem cell transplantation groups, the left ventricular systolic pressure and maximum rise/fall rate of left ventricular pressure were significantly decreased, but the left ventricular end diastolic pressure significantly increased in the model group (P < 0.05). And there was no significant difference between the normal group and the stem cell transplantation group (P > 0.05). High and dense fluorescence intensity was observed in the host myocardium immediately after transplantation. Subsequently, the fluorescence intensity and density decreased at 4 weeks, but the cell migration could be found, and some cells expressed cardiac troponin T. These results show that intravenous transplantation of bone marrow mesenchymal stem cells can improve cardiac function in rats with heart failure due to cardiomyopathies. Besides, the transplanted cells can survive in the host, and differentiate into cardiomyocyte-like cells.     

电针刺激对神经干细胞移植治疗脊髓损伤大鼠后肢功能的影响

背景：单纯的神经干细胞移植对受损脊髓组织的修复作用并不理想,为了进一步提高移植细胞在体内的存活、增殖及定向分化为神经元的比例,必须进一步改善脊髓损伤区的微环境。 目的：观察神经干细胞移植联合电针刺激对脊髓损伤大鼠后肢功能及电生理的影响。 方法：将脊髓损伤模型SD大鼠72只按随机数字表法分为4组：对照组尾静脉注入培养液,神经干细胞组经尾静脉注入等体积神经干细胞悬液,电针刺激组自模型完成6 h起采用督脉加体穴电针1周,联合组尾静脉注射神经干细胞后,同时采用督脉加体穴电针1周。分别于造模前、造模后1,3 d、1-4 周通过BBB评分、斜板试验进行运动功能评定。造模后4周取材行病理切片苏木精-伊红染色,荧光显微镜观测CM-Dil 标记的神经干细胞存活及分布情况,辣根过氧化物酶示踪观察神经纤维再生情况,运动诱发电位和体感诱发电位观察大鼠神经电生理恢复情况。 结果与结论：造模后2-4周大鼠下肢运动功能评价联合组优于神经干细胞组及电针刺激组,神经干细胞组和电针刺激组优于对照组。造模后4周,神经干细胞组和电针刺激组损伤区可见少量神经轴索样结构,脊髓空洞较小,联合组可见较多神经轴索样结构,未见脊髓空洞。造模后4周,CM-Dil 阳性细胞和辣根过氧化物酶阳性神经纤维数：联合组>神经干细胞组与电针刺激组>对照组,各组之间差异有显著性意义(P < 0.05)。运动诱发电位和体感诱发电位的潜伏期：联合组<神经干细胞组与电针刺激组<对照组,各组之间差异有显著性意义(P < 0.05);运动诱发电位和体感诱发电位的波幅：联合组>神经干细胞组与电针刺激组>对照组,各组之间差异有显著性意义(P < 0.05)。结果提示神经干细胞移植的同时联合电针刺激能够促进脊髓损伤大鼠神经突触的再生,改善其大鼠肢体运动功能及电生理功能。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程      

舒芬太尼复合丙泊酚全身麻醉在脐血间充质干细胞移植治疗脊髓损伤中的应用

BACKGROUND:Sufentanil and propofol are both found to have good neuroprotective effects on neurological damage in clinical practice. OBJECTIVE:To investigate the effect of propofol combined with sufentanil in umbilical cord blood mesenchymal stem cells transplantation for the treatment of spinal cord injury. METHODS:Sixty-five Wistar rats were selected to make animal models of acute spinal cord injury using Allen’s method. Six hours after modeling, these rats were randomly assigned into combined group (injection of 2×10⁷/L human umbilical cord blood mesenchymal stem cell suspension (0.5 mL) plus injection of 1.0-1.5 mg/kg propofol and 0.5 μg/kg sufentanil via the tail vein), stem cell group (injection of 2×10⁷/L human umbilical cord blood mesenchymal stem cell suspension (0.5 mL) via the tail vein), or control group (injection of 30 μL of LDMEM containing 5% fetal bovine serum). S100β protein level in serum was detected in each group at 15 and 60 minutes after injection. Motor function of rat in each group was assessed by Basso Beattie Bresnahan (BBB) scoring and incline plane test at 1, 2, 4, 6, 8 weeks after modeling. Pathological changes of the spinal cord were observed at 4 weeks after modeling. Expression of vascular endothelial growth factor was detected using western blot assay at 1 and 2 weeks after modeling. RESULTS AND CONCLUSION:After 15 and 60 minutes of intervention, S100β protein level was lowest in the combined group followed by the stem cell and control groups (P < 0.05). At 2, 4, 6, 8 weeks after modeling, scores on the incline plane test and BBB were ranked as follows: combined group > stem cell group > model group (P < 0.05). At 4 weeks after modeling, severe damage to the spinal cord and few nerve fibers were found in the control group; spinal cord hyperplasia and a few of regenerated axons and PKH-26-positive stem cells appeared in the stem cell group; while in the combined group, there were a large amount of PKH-26-positive stem cells and nerve axon-like structures. At 1 and 2 weeks after modeling, the highest protein level of vascular endothelial growth factor was found in the combined group followed by the stem cell group and control group (P < 0.05). To conclude, these findings indicate that propofol and sufentanil in umbilical cord blood mesenchymal stem cell transplantation therapy can promote the recovery of hindlimb function after spinal cord injury, thereby promoting the functional recovery of rats from spinal cord injury.     

乌司他丁联合脐带间充质干细胞移植脊髓损伤大鼠后肢运动功能和 运动诱发电位的变化

背景：乌司他丁能减轻炎性反应、清除氧自由基,对中枢神经系统损伤具有保护作用,能有效地提高脊髓损伤后移植细胞的存活率。 目的：观察乌司他丁联合脐带间充质干细胞移植对脊髓损伤大鼠后肢功能的影响。 方法：Wistar大鼠建立脊髓损伤动物模型后随机分成4组：空白对照组尾静脉注射培养液+腹腔注射生理盐水,细胞移植组尾静脉注射脐带间充质干细胞,乌司他丁组腹腔注入乌司他丁,联合组尾静脉注射脐带间充质干细胞,同时腹腔注入乌司他丁。 结果与结论：移植4周后联合组下肢运动功能优于细胞移植组和乌司他丁组(P < 0.05),细胞移植组和乌司他丁组优于空白对照组(P < 0.05)。移植后4周,PKH26标记的阳性细胞数联合移植组多于细胞移植组,细胞移植组多于乌司他丁组和空白对照组(P < 0.01)。移植后8周,联合组大鼠体感诱发电位及运动诱发电位的潜伏期、波幅明显优于其他3组(P < 0.05或P < 0.01)。提示乌司他丁联合应用脐带间充质干细胞移植可促进脊髓损伤大鼠神经突触的再生,改善其肢体运动功能和电生理功能,其效果优于单独应用乌司他丁或脐带间充质干细胞。     

低频电磁场促进骨髓间充质干细胞移植修复大鼠脊髓损伤的实验研究

背景：骨髓间充质干细胞移植治疗脊髓损伤被视为一种有前途的治疗方法,如何更有效地促进骨髓间充质干细胞在脊髓损伤区存活,加速脊髓损伤肢体运动功能的恢复是目前研究的重点。前期研究发现,低频电磁场能够促进骨髓间充质干细胞的增殖分化,低频电磁场是否可应用于骨髓间充质干细胞移植治疗脊髓损伤还需进一步研究。 目的：探讨低频电磁场对移植骨髓间充质干细胞脊髓损伤大鼠后肢运动功能恢复的影响。 方法：采用脊髓压迫法制备64只T10不完全性脊髓损伤大鼠模型,随机等分为对照组、骨髓间充质干细胞组、电磁场组和电磁场+骨髓间充质干细胞组。造模成功后,骨髓间充质干细胞组和电磁场+骨髓间充质干细胞组大鼠脊髓损伤原部位注射大鼠全贴壁法分离培养BrdU标记的骨髓间充质干细胞,对照组和电磁场组注射a-MEM培养液。造模术后24 h,电磁场组和电磁场+骨髓间充质干细胞组予60 min/d的低频电磁场刺激(频率50 Hz、强度5 mT)。 结果与结论：骨髓间充质干细胞移植后第21天,电磁场+骨髓间充质干细胞组BBB评分与其他组相比,差异有显著性意义(P < 0.05),与其他各组比较,电磁场+骨髓间充质干细胞组移植细胞后,大鼠BrdU阳性细胞在脊髓损伤区域生长并与脊髓组织融合,存活细胞数量较其他组多;空洞面积小;损伤区胶质纤维酸性蛋白表达更少,而基质金属蛋白2表达更多;脊髓损伤大鼠下肢运动功能恢复最快(P < 0.05)。提示低频电磁场促进了移植骨髓间充质干细胞脊髓损伤大鼠后肢运动功能的恢复,可能与低频电磁场有利于损伤区移植骨髓间充质干细胞的存活,上调基质金属蛋白2的表达并减少胶质瘢痕的形成有关。     

依达拉奉联合神经干细胞移植修复大鼠脊髓损伤

背景：依达拉奉是一种自由基清除药,可以减轻受损神经组织水肿和改善脊髓损伤区微环境。 目的：观察依达拉奉联合神经干细胞移植对大鼠脊髓全横断损伤的修复效果。 方法：成年雌性SD大鼠80只,建立胸9脊髓全横断损伤模型,随机分为4组：对照组不做处理;依达拉奉组脊髓损伤后6 h经尾静脉注射依达拉奉;神经干细胞移植组脊髓损伤后6 h脊髓损伤区域注入神经干细胞悬液;依达拉奉+细胞移植组脊髓损伤后6 h神经干细胞移植的同时尾静脉注射依达拉奉。 结果与结论：造模后8周可观察到PKH-26标记的神经干细胞在体内存活并在脊髓内迁移;细胞移植组和依达拉奉联+细胞移植组可见少量连续性神经纤维通过损伤区。荧光金逆行脊髓追踪显示神经干细胞移植组和依达拉奉+细胞移植组可见被荧光金标记的神经锥体细胞穿越损伤区。PKH-26标记的阳性细胞数及荧光金阳性神经纤维数：依达拉奉+细胞移植组最多,依达拉奉组、神经干细胞移植组次之,对照组最少,各组之间差异有显著性意义(P < 0.05);后肢功能运动BBB评分依次为依达拉奉+细胞移植组>神经干细胞移植组>依达拉奉组>对照组。提示依达拉奉能促进神经干细胞在损伤区的存活并向神经细胞分化,依达拉奉联合神经干细胞移植有促进细胞移植修复大鼠脊髓损伤的效果。     

内皮祖细胞移植联合早期运动改善脊髓损伤区血管再生及后肢功能

BACKGROUND: Endothelial progenitor cells are widely used in the treatment of various vascular diseases, and early exercise training contributes to restore motor function after spinal cord injury. However, the therapeutic effects of endothelial progenitor cell transplantation or early exercise training alone are unfavorable. OBJECTIVE: To observe the influence of transplantation of endothelial progenitor cells combined with early exercise training on blood vessel regeneration and hind limb function in rats after spinal cord injury. METHODS: Eighty adult Sprague-Dawley rats were enrolled to establish spinal cord injury models using the modified Allen’s method, and then randomly divided into four groups. Rats were respectively given culture medium via the tail vein, injection of endothelial progenitor cells (3×106) via the tail vein, roller and treadmill trainings for 2 weeks, or injection of endothelial progenitor cells via the tail vein followed by 2 weeks of roller and treadmill trainings in the model, cell transplantation, exercise and combined groups. RESULTS AND CONCLUSION: At 2 weeks after transplantation, the hindlimb motor function of rats in the combined group was better than that in the cell transplantation group and exercise group, and moreover, the percentage of CM-Dil positive cells, the number of horseradish peroxidase-positive nerve fibers, capillary density and expression of vascular endothelial growth factor and brain-derived neurotrophic factor were also significantly higher in the combined group than the cell transplantation group and exercise group. These findings indicate that early exercise training has a neuroprotective role in spinal cord injury; endothelial progenitor cell transplantation combined with early exercise training can promote regeneration of synapses and blood vessels and improve hindlimb motor function of rats, probably by increasing expression levels of vascular endothelial growth factor and brain-derived neurotrophic factor.       

白细胞介素6受体单克隆抗体与骨髓间充质干细胞可减少急性脊髓损伤神经元的凋亡

 BACKGROUND: Studies have suggested that interleukin-6 is crucial for inducing cell apoptosis after acute spinal cord injury. OBJECTIVE: To observe the effect of interleukin-6 receptor monoclonal antibody combined with bone marrow mesenchymal stem cells to treat acute spinal cord injury in rats. METHODS: Thirty Sprague-Dawley rats were randomly divided into sham group, model group (spinal cord injury group), treatment group 1 (interleukin-6 receptor monoclonal antibody transplantation group), treatment group 2 (bone marrow mesenchymal stem cell transplantation group), treatment group 3 (bone marrow mesenchymal stem cell+interleukin-6 receptor monoclonal antibody group), with six rats in each group. In the sham group, the spinal cord was only exposed with no injury, and in the other four groups, rat models of acute spinal cord injury were made using modified Allen’s method. Local injection treatment was performed in all the groups at 28 days after modeling. Basso, Beattie and Bresnahan (BBB) scoring and improved Tarlov scoring were used at 1 day before treatment and 1, 3, 7, 14, 28 days after treatment to test the hindlimb function. At 28 days after treatment, TUNEL method was used to detect cell apoptosis in the spinal cord. RESULTS AND CONCLUSION: Compared with the sham group, BBB scores and improved Tarlov scores were decreased significantly in the other four groups (P < 0.05). At 7 days after treatment, the BBB scores and improved Tarlov scores in the treatment group 3 were significantly higher than those in the model group (P < 0.05). At 14 days after treatment, the BBB scores and improved Tarlov scores in the treatment groups 1 and 2 were significantly higher than those in the model group (P < 0.05); compared with the treatment group 2, the BBB score and improved Tarlov score were significantly increased in the treatment group 3 (P < 0.05). Compared with the sham group, the number of apoptotic cells was significantly increased in the other four groups (P < 0.05); compared with the model group, the number of apoptotic cells was significantly decreased in the three treatment groups (P < 0.05); compared with the treatment group 2, the number of apoptotic cells was significantly lower in the treatment group 3 (P < 0.05). These findings indicate that the combined use of interleukin-6 receptor monoclonal antibody and bone marrow mesenchymal stem cell transplantation is better than bone marrow mesenchymal stem cell transplantation alone in the treatment of spinal cord injury, and interleukin-6 receptor monoclonal antibody reduces cell apoptosis in spinal cord injury, which is of positive significance for preventing against acute spinal cord injury.     

骨髓间充质干细胞移植脊髓损伤小鼠运动功能的恢复

背景：研究证实,移植的骨髓间充质干细胞可被定向诱导分化为神经细胞,重建神经环路,促进轴突再生,恢复脊髓功能。 目的：进一步验证骨髓间充质干细胞在脊髓损伤修复中的作用。 方法：C57BL/6小鼠40只随机分为4组,假手术组不打击脊髓;其余小鼠采用重物撞击法建立脊髓损伤模型。损伤后的第7天,治疗组用微量注射器,经眶静脉丛注入骨髓间充质干细胞悬液;对照组注入等量 DMEM培养基;模型组不做处理。通过苏木精-伊红染色法判断脊髓损伤程度。通过免疫细胞化学法鉴定骨髓间充质干细胞分化形成的神经细胞。通过荧光显微镜观察移植细胞生长状态;通过改良Tarlov评分法评价小鼠运动功能恢复程度。 结果与结论：骨髓间充质干细胞诱导分化7 d后的细胞呈NF和神经胶质纤维酸性蛋白阳性表达。模型组小鼠双下肢呈瘫痪状态,假手术组行动正常(P < 0.01)。细胞移植后2周,治疗组小鼠运动功能缺失症状逐渐恢复,对照组小鼠恢复不明显 (P < 0.05);细胞移植4周后,细胞移植组小鼠Tarlov评分与假手术组相比差异无显著性意义(P > 0.05)。说明骨髓间充质干细胞移植可提高脊髓损伤小鼠的运动能力。     

骨髓间充质干细胞向脊髓损伤区的迁徙

背景：骨髓间充质干细胞移植到脊髓损伤区域后,如何观察其在体内的生存和转归情况,一直是让人困扰的问题。 目的：观察骨髓间充质干细胞在大鼠脊髓损伤区内的迁徙情况。 方法：36只Wistar大鼠随机分为2组,实验组制作脊髓损伤模型1周后,经尾静脉移植用DAPI标记的骨髓间充质干细胞(1×109 L-1) 1 mL,连续注射2 d。对照组未行脊髓损伤,与实验组同一时间同法行骨髓间充质干细胞移植。分别于移植后5,10,15 d,制作损伤脊髓冰冻切片,在激光共聚焦显微镜下观察骨髓间充质干细胞的迁徙情况。 结果与结论：实验组于移植后5 d,在脊髓损伤组织血管内出现少量荧光标记的骨髓间充质干细胞,10 d后有血管外弥散,15 d后有广泛弥散。对照组均未见DAPI标记的骨髓间充质干细胞。结果表明骨髓间充质干细胞经大鼠尾静脉移植后,能透过血脊髓屏障向损伤脊髓组织迁徙。     

骨髓间充质干细胞移植脊髓全横断模型大鼠运动和体感诱发电位的评价

背景：临床常用皮质运动诱发电位和皮质体感诱发电位来分别评价脊髓损伤后运动传导路和感觉传导路的损伤或修复情况。 目的：以脊髓诱导电位监测骨髓间充质干细胞移植后急性脊髓完全性损伤大鼠下肢神经功能的变化。 方法：选取健康Wistar大鼠50只,分成5组,即生理盐水组、骨髓间充质干细胞移植组、脑源性神经营养因子修饰组、神经营养素3+骨髓间充质干细胞移植组和假手术组。除假手术组外,其余各组均制作Allen’s脊髓完全性损伤动物模型,造模后各组均行相应治疗。治疗后4,8和12周行大鼠后肢运动功能评分,并于造模后24 h,3,7,14 d行运动和体感诱发电位检测。 结果与结论：运动诱发电位检测结果提示,各治疗组的运动功能均有不同程度的恢复,与生理盐水组间差异均有显著性意义(P < 0.05),大鼠后肢BBB评分也证实了各治疗组后肢运动功能明显优于生理盐水组(P < 0.05)。提示经脑源性神经营养因子修饰的骨髓间充质干细胞可移植到脊髓损伤处,可改善大鼠的后肢运动,神经营养素3蛋白有可能提高骨髓间充质干细胞在体内的生存率,促进受损脊髓的轴突再生。