中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (45): 8431-8434.doi: 10.3969/j.issn.1673-8225.2010.45.017

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

Brca-1基因调控神经干细胞移植治疗脑梗死

方力群，张卓伯   

  1. 哈尔滨医科大学附属第四医院神经内科，黑龙江省哈尔滨市  150001
  • 出版日期:2010-11-05 发布日期:2010-11-05
  • 通讯作者: 张卓伯，博士，副主任医师，哈尔滨医科大学附属第四医院神经内科，黑龙江省哈尔滨市 150001
  • 作者简介:方力群☆，女，1970年生，黑龙江省哈尔滨市人，汉族，在读博士，副主任医师，主要从事脑血管病的治疗及神经心理方面的研究。 fangliquncongcong@yahoo.com.cn

Brca-1 gene-regulated neural stem cells transplantation in treatment of cerebral infarction

Fang Li-qun, Zhang Zhuo-bo   

  1. Department of Neurology, Fourth Affiliated Hospital, Harbin Medical University, Harbin  150001, Heilongjiang Province, China
  • Online:2010-11-05 Published:2010-11-05
  • Contact: Zhang Zhuo-bo, Doctor, Associate chief physician, Department of Neurology, Fourth Affiliated Hospital, Harbin Medical University, Harbin 150001, Heilongjiang Province, China
  • About author:Fang Li-qun☆, Studying for doctorate, Associate chief physician, Department of Neurology, Fourth Affiliated Hospital, Harbin Medical University, Harbin 150001, Heilongjiang Province, China fangliquncongcong@yahoo.com.cn

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被引次数

2

摘要：

背景：最近国际上有人推测神经前体细胞和神经细胞的增殖受到Brca-1基因调控。
目的：观察Brca-1基因调控神经干细胞移植对SD大鼠急性缺血性脑梗死模型影响。
方法：参照Longa等线栓法建立SD大鼠急性脑梗死模型。应用立体定位法，分别向模型大鼠病侧脑室内注射PBS溶液、常规培养的神经干细胞、10-5 mol/L 17β-雌二醇培养的神经干细胞。采用动物行为学评分评价模型动物不同时间点的神经功能缺损；激光共聚焦显微镜观察神经干细胞Nestin表达；TTC染色测定不同时间点脑梗死体积变化；光镜和电镜分别观察脑梗死区病理变化。免疫组织化学法检测NSE和GAFP表达。
结果与结论： ①17β-雌二醇培养组动物神经功能恢复程度优于常规培养组(P < 0.05)。②激光共聚焦显微镜显示脑缺血再灌注组和空白干预组在4 d时荧光强度最强，7 d时荧光表达减弱，17β-雌二醇培养组也是在4 d时表达最强，但是持续表达高强度的时间长，7 d时仍能见到较强的荧光表达。③17β-雌二醇培养组脑梗死体积明显缩小，与脑缺血再灌注组和空白干预组相比差异有显著性意义(P < 0.05)。④17β-雌二醇培养组细胞移植治疗7 d神经元基本无肿胀，核仁清晰，染色质分布均匀，大部分毛细血管形态正常。⑤17β-雌二醇培养组NSE和GFAP表达的阳性结果也高于常规培养组。结果表明用雌激素干预Brca-1基因表达的神经干细胞移植后治疗效果好于常规培养的神经干细胞。

关键词: 雌二醇, 脑梗死, 动物模型, 细胞移植, Brca-1, 神经干细胞

Abstract:

BACKGROUND: Recently, someone presumed that the proliferation of neural precursor cells and neural cells is regulated by Brca-1 gene.
OBJECTIVE: To observe the effect of Brca-1 gene regulation of neural stem cell transplantation on Sprague Dawley rat models of acute ischemic cerebral infarction.
METHODS: Sprague Dawley rat models of acute ischemic cerebral infarction were established according to Longa method. By stereotaxical method, phosphate buffered saline, routinely cultured neural stem cells and 10-5 mol/L 17 β-estradiol cultured neural stem cells were infused into the lateral ventricle of affected brain of rat models separately. Neurological impairment was evaluated at various time points using animal ethology score. Laser confocal microscopy was utilized to observe Nestin expression in neural stem cells. Cerebral infarction volume on cerebral ischemia reperfusion model rats was evaluated by 2, 3, 5-rihenyltetrazolium chloride (TTC) staining. The brain pathological changes were observed by the optical and electron microscope. Immunohistichemical staining was used to examine the expression of neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP).
RESULTS AND CONCLUSION:  ① The recovery of neural function was better in the 17 β-estradiol group compared with routinely cultured group (P < 0.05). ② Laser confocal microscopy has revealed that fluorescence intensity was strongest at 4 days in the ischemia/reperfusion group and blank group, gradually weak at 7 day. The fluorescence intensity was strongest at 4 days in the 17 β-estradiol group, but the persistent high intensity was long, and the strong intensity was visible at 7 days. ③ The cerebral infarct volume was significantly shortened in the 17 β-estradiol group, and significant difference was detected compared with the ischemia/reperfusion and blank groups (P < 0.05). ④ Neurons were not swollen, with visible nuclei, even chromatin and a majority of normal blood capillary at 7 days following cell transplantation in the 17 β-estradiol group. ⑤ Positive expression of NSE and GFAP was greater in the 17 β-estradiol group compared with routinely cultured group. These suggest that the neuroprotective effect of neural stem cells cultured with estrogen transplantation group is better than routine neural stem cell transplantation group.

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