自体脂肪干细胞移植脑冻伤大鼠脑内的作用  

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

作　　者：周向阳[1] 邓永文[1] 方芳[2] 王飞[1] 陈若琨[1] 宋涛[1] 方加胜[1] 

机构地区：[1]中南大学湘雅医院神经外科,湖南省长沙市410008 [2]中南大学湘雅医院心内科,湖南省长沙市410008

出　　处：《中国组织工程研究与临床康复》2008年第16期3024-3028,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

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300g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1,3,5,7,14,30d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3-14dNSS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,3d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。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. （1）SD 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. （2）ADSCs 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. （3） Neurological 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： （1）The decrease in NSS were found in rats of the experimental group in 3-14 days （P 〈 0.05）. （2）BrdU labeled stem cells survived in rat brain tissue and can be transferred detected by immunofluorescence. （3）The apoptosis was obviously inhibited in the experimental group through Tunel investigation 3 days later. （4）The expressions of VEGF and BDNF wer

关 键 词：脂肪干细胞 脑冷冻伤 凋亡 移植 神经行为学 

分 类 号：R394.2[医药卫生—医学遗传学]