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脐血间充质干细胞静脉移植治疗缺氧缺血性脑损伤的时效性(英文) 被引量:5

Chronergy of umbilical cord blood-mesenchymal stem cell transplantation for treatment of hypoxic-ischemic brain damage
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摘要 背景:研究证实,使用间充质干细胞移植治疗成年动物缺氧缺血性脑损伤已取得了一定的疗效,但新生动物模型较少见。治疗所使用的间充质干细胞基本上都来源于骨髓,使用脐血间充质干细胞的研究报道较少。目的:将脐血间充质干细胞从尾静脉注入新生鼠缺氧缺血性脑损伤模型,探讨脐血间充质干细胞移植治疗的可行性及时效性。设计、时间和地点:完全随机对照的动物实验,于2004—10/2005—07在南昌大学第一附属医院神经内科实验室完成。材料:选用健康新生7d龄SD大鼠38只制作新生大鼠缺氧缺血性脑损伤模型,死亡3只。方法:采集23~35岁健康孕妇足月顺产儿的脐带血体外培养脐血间充质干细胞。移植前应用4’,6二脒基-2-苯吲哚盐酸体外标记间充质干细胞。造模成功的35只大鼠随机分为3组:早期移植组12只造模后第2天经鼠尾静脉注入脐血间充质干细胞;晚期移植组12只造模后1周进行移植;对照组11只注射等量的生理盐水。早期移植组和晚期移植组于移植后第2天以及造模后2周分别随机取6只大鼠,对照组于造模后2d,1和2周分别取3,4,4只大鼠麻醉后处死。取脑和海马回区的缺血脑组织制作冰冻切片。主要观察指标:苏木精-伊红染色观察脑组织病理形态学改变。荧光显微镜观察脑组织4’,6二脒基-2-苯吲哚盐酸阳性细胞。结果:对照组缺血区脑水肿,神经细胞肿胀,细胞数减少;早期移植组于移植后病灶侧脑内很少见到4’,6二脒基-2-苯吲哚盐酸阳性脐血间充质干细胞分布,其脑组织水肿程度及细胞外间隙的改善和细胞数目的增加也不明显;晚期移植组,造模后1周缺血脑组织细胞外间隙缩小,细胞数明显增加,脑组织水肿已明显减轻,在大鼠病灶侧脑内,可见大量的4’,6二脒基-2-苯吲哚盐酸阳性细胞向病灶区及周围迁移和扩散,没有明显的界限。结论:脐血间充质干细胞能有效透过血脑屏障,在病灶脑组织周围迁移、扩散、整合;选择缺氧缺血性脑损伤后1周时进行移植效果较好。 BACKGROUND: Mesenchymal stem cell transplantation for treatment of hypoxic-ischemic brain damage (HIBD) has obtained some outcomes in adult animals, but studies are few in neonatal animal models. Mesenchymal stem cells are commonly harvested from bone marrow. A few studies are on umbilical cord blood-mesenchymal stem cells (UCB-MSCs). OBJECTIVE: To investigate the feasibility and timeliness of UCB-MSC transplantation after injecting UCB-MSCs into neonatal rat models of HIBD. DESIGN, TIME AND SETTING: The complete randomized controlled animal experiment was performed at the Laboratory of Department of Neurology of First Hospital Affiliated to Nanchang University from October 2004 to July 2005. MATERIALS: A total of 38 healthy neonatal SD rats aged 7 days old were used to create rat models of HIBD. Three rats died. METHODS: Cord blood samples were collected after normal full-term delivery of 23-35 healthy pregnant women for culturing UCB-MSCs. MSCs were labeled with 4', 6-diarnidino-2-phenylindole 2hci (DAd?I) in vitro before transplantation. Thirty-five rat models were divided into three groups. UCB-MSCs were injected into tail vein of twelve rat models in the early transplantation group two days after modeling. UCB-MSCs were injected into tail vein of twelve rat models in the late transplantation group one week after modeling. Same volume of saline was injected into eleven rats of the control group. Six rats from early transplantation and late transplantation groups each were respectively obtained at day 2 after transplantation and at week 2 after modeling. Three, four and four rats from control group were obtained respectively 2 days, 1 and 2 weeks after modeling, and sacrificed after anaesthesia. Ischemic brain tissues from the brain and hippocampal gyrum were sliced into frozen sections. MAIN OUTCOME MEASURES: Brain tissue pathomorphology was measured by Haematoxylin and Eosin Staining. Brain tissue DAPI-positive cells were detected with a fluorescence microscope. RESULTS: Brain edema at ischemic region, neural cell swelling and a decrease in cell number were tested in the control group. DAPI-positive UCB-MSCs were few in focal brain tissues, and swelling degree, extracellular space improvement and increased cell number were insignificant in the early transplantation group. One week after modeling, brain tissue extracellular space became small, cell number increased, and brain swelling reduced; A mass of DAPI-positive cells in rat focal brain migrated and diffused, without significant boundary in the late transplantation group. CONCLUSION: UCB-MSCs effectively traverse blood-brain barrier, and migrate, disperse and conform around focal brain tissues. A good outcome of transplantation is obtained at week 1 after HIBD.
出处 《中国组织工程研究与临床康复》 CAS CSCD 北大核心 2008年第25期4975-4978,共4页 Journal of Clinical Rehabilitative Tissue Engineering Research
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  • 1吴婉芳,徐放生,张莉莉,蔡玲玲,林久治.建立新生儿缺氧缺血性脑病动物模型[J].中国新生儿科杂志,1992,17(6):265-267. 被引量:128
  • 2赵晓科,肖农,周江堡,张晓萍.麻黄碱对脑缺血大鼠运动功能恢复的影响及分子机制研究[J].中国康复医学杂志,2005,20(3):172-175. 被引量:12
  • 3鞠秀丽,黄志伟,时庆,侯怀水,段春红.体外扩增脐血间充质干细胞的生物学特性和诱导分化能力的研究[J].中华儿科杂志,2005,43(7):499-502. 被引量:17
  • 4Rice JE, Vannucci RC, Brierley JB, et al. The influence of immaturity on hypoxic-ischemic brain damage in the rat [ J]. Ann Neurol, 1981, 9:131-141.
  • 5Andine P, Thordstein M, Kjellmer I, et al. Evaluation of brain damage in a rat model of neonatal hypoxic-ischemla [J]. J Neurosci Methods, 1990, 35:253-260.
  • 6Vannucci RC, Vannucci SJ. A model of perinatal hypoxic-ischemic brain damage [ J ]. Ann N Y Acad Sci ,1997,835:234-249.
  • 7Wagner CL, Either DJ, Katikaneni LD, et al. The use of hypothermia: a role in the treatment of neonatal asphyxia [J] ? Pediatr Neurol, 1999,21:429-443.
  • 8Thoresen M. Cooling the asphyxiated brain-ready for clinical trials[J] .Eur J Pediatr, 1999, 158 (suppl 1 ):55-58.
  • 9Waters CM, Moser W, Walkinshaw G, et al. Death of neurons in the neonatal rodent and primate globus pallidus occurs by a mechanism of apoptosis [ J]. Neuroscience, 1994, 63:881-894.
  • 10Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science, 1999, 284:143-147.

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