相关链接：药物治疗神经元功能缺损引起的一些严重的疾病时遇到苦难，因此学者们已经将目光投向干细胞这一新兴领域，干细胞具有增殖并且向多种细胞系列分化的能力。当前干细胞的研究主要集中在诱导分化、细胞移植以及以干细胞为基因载体的的研究。目前，干细胞在神经退行性疾病的研究方面已经取得一些成果，比如其对帕金森等疾病的治疗已经进入临床。由于定向诱导、干细胞迁移、损伤后炎症以及胶质瘢痕增生以及神经元突触联系等问题都还未解决，干细胞对于神经系统损伤的治疗一方面却进展相对缓慢，这些问题也是我们今后研究的重点。

应用要点：已有实验表明，通过一些诱导方式脂肪干细胞完全可以向神经系细胞分化。因此实验选用脂肪干细胞作为移植细胞，制作便于控制温度以及时间的冷冻伤模型，很好的模拟了脑外伤以后的原发和继发性脑损伤。脑损伤后移植脂肪干细胞，结果发现其可以很好的在宿主体内存活、分化并且迁移到达受损病灶部位，并发现血管内皮生长因子、脑源性神经营养因子等基因的表达上调。

同行评价：有研究表明，干细胞具有在体内外诱导分化成神经元样细胞的潜能。干细胞移植修复神经损伤，成为近年来神经科学领域中倍受学者关注的研究热点之一。本文通过制作大鼠脑冷冻伤模型，采用体外培养并传代SD大鼠脂肪干细胞并移植入大鼠脑内，检测BrdU阳性细胞和凋亡细胞及血管内皮生长因子、脑源性神经营养因子表达，选题较新颖，有一定学术创新和临床应用意义。

摘要
目的：脂肪干细胞来源于成脂细胞，取材容易，已经成为干细胞研究一个新的方向。观察脂肪干细胞移植脑冻伤大鼠的脑组织局部组织学变化以及神经行为学变化。
方法：实验于2006-01/2007-08在中南大学湘雅医院神经病学实验室完成。①实验材料：SD大鼠，雌雄不限，体质量300 g左右，由中南大学湘雅医学院动物学部提供。实验过程中对动物处置符合动物伦理学标准。②实验方法：体外培养并传代SD大鼠脂肪干细胞,并用BrdU标记。制作大鼠的脑冷冻伤模型，实验组在冷冻伤动物脑内移植脂肪干细胞，对照组在冷冻伤动物脑内移植培养基，假手术组仅行颅骨开窗，每组15只。③实验评估：分别在移植细胞后1，3，5，7，14，30 d对实验动物进行NSS神经行为学评分，并制作脑组织切片，行免疫荧光染色检测BrdU阳性细胞。采用Tunel染色检测凋亡细胞，脑组织RT-PCR检测血管内皮生长因子、脑源性神经营养因子等因子mRNA表达。
结果：①与对照组相比，实验组大鼠在3~14 d NSS神经行为学评分降低（P < 0.05）。②免疫荧光检测显示，Brdu标记的脂肪干细胞在大鼠脑组织内存活并且迁移。③Tunel法检测显示，3 d后实验组凋亡细胞数目明显少于其他组。④RT-PCR检测显示，与其他两组比较，移植脂肪干细胞的大鼠脑组织中血管内皮生长因子、脑源性神经营养因子表达更高。
结论：脂肪干细胞可以在中枢神经系统中存活，并分化为神经元样细胞，它可以引起血管内皮生长因子、脑源性神经营养因子等因子的高表达从而减少细胞凋亡，加速神经功能修复过程并达到保护脑组织的目的。
关键词：脂肪干细胞；脑冷冻伤；凋亡; 移植；神经行为学

周向阳，邓永文，方芳，王飞，陈若琨，宋涛，方加胜.自体脂肪干细胞移植脑冻伤大鼠脑内的作用[J].中国组织工程研究与临床康复，2008，12(16):3024-3028 [www.zglckf.com/zglckf/ejournal/upfiles/08-16/16k-3024(ps).pdf]

Autologous adipose tissue-derived stem cell transplantation for treatment of rats with brain cold injury

Abstract

AIM：Adipose tissue-derived stem cells (ADSCs) can be easily separated from the fat tissue and now become a new direction of the research on stem cells. This study aimed to investigate the histologic and neurological changes in SD rats with brain cold injury after transplanting the ADSCs into the brain.
METHODS: Experiments were performed at the Neurology Lab of Xiangya Hospital of Central South University from January 2006 to August 2007. ①SD rats of about 300 g of either gender were provided by the Experimental Animal’s Department of Xiangya Medical College of Central South University. The disposal to the animals was accorded with the ethical standards. ②ADSCs from SD rats were cultured and labeled with BrdU in vitro, and then transplanted into the SD rats with brain cold injury were induced previously. Animals were divided into 3 groups, an experimental group (ADSCs was transplanted), a control group (medium was transplanted) and a sham operation group (only craniotomy was executed without transplantation) with 15 rats in each group. ③Neurological Severity Scores (NSS) were evaluated at 1, 3, 5, 7, 14 and 30 days after cell transplantation to the experimental animals. The brain tissues of the animals were sliced after being executed death. Immunofluorescence was used to detect the BrdU positive cells. Frozen brain sections were subjected to Tunel staining to investigate the apoptosis cells. Reverse transcription-polymerase chain reaction (RT-PCR) was used to find the mRNA expression of vascular endothelial growth factor (VEGF) and brain-derived neurotrophic factor (BDNF).
RESULTS: ①The decrease in NSS were found in rats of the experimental group in 3-14 days (P < 0.05). ②BrdU labeled stem cells survived in rat brain tissue and can be transferred detected by immunofluorescence. ③The apoptosis was obviously inhibited in the experimental group through Tunel investigation 3 days later. ④The expressions of VEGF and BDNF were higher in the brain tissue of the experimental group than the control and sham operation groups detected by RT-PCR.
CONCLUSION: ADSCs can survive and differentiate into neuron-like cells in the central nervous system. ADSCs transplantation can reduce apoptosis by evoking the high expression of factors like BDNF and VEGF, through which neural regeneration is accelerated and then brain is protected in injury finally.

Zhou XY, Deng YW, Fang F, Wang F, Chen RK, Song T, Fang JS.Autologous adipose tissue-derived stem cell transplantation for treatment of rats with brain cold injury.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(16):3024-3028
[www.zglckf.com/zglckf/ejournal/upfiles/08-16/16k-3024(ps).pdf]