摘要
目的研究胎鼠神经干细胞(NSCs)移植对大鼠脊髓损伤(SCI)后神经细胞凋亡及凋亡抑制基因Bcl-2表达的影响。方法 40只SD大鼠随机分为正常对照组(Normal组),脊髓损伤组(SCI组),神经干细胞组(NSC组),神经干细胞标记组(BrdU+NSCs组)。采用电控脊髓损伤打击装置制作模型,5-溴脱氧尿嘧啶核苷(Br-dU)法标记处于对数生长期的NSCs,SCI后即刻进行NSCs移植。免疫组化法观察BrdU标记NSCs的存活、迁移及凋亡抑制基因Bcl-2的表达,TUNEL法标记凋亡细胞(免疫组化及免疫荧光显色),改良Rivlin法观察大鼠后肢运动功能的恢复情况。结果 BrdU+NSCs组在损伤脊髓区域可检测到BrdU标记的阳性NSCs。BrdU+NSC组与NSC组各时间点凋亡阳性细胞数均比SCI组减少(P<0.01),Bcl-2免疫阳性细胞光密度值比SCI组明显增加(P<0.01),且Bcl-2表达高峰延长至伤后7d;移植后7d、14d、28d后肢运动功能评分较SCI组明显升高(P<0.01)。Br-dU+NSC组与NSC组之间比较无明显差异(P>0.05)。结论体外培养的胚胎大鼠NSCs可在脊髓损伤区域存活、迁移,并能通过上调Bcl-2的表达来抑制大鼠脊髓损伤后神经细胞的凋亡,从而促进大鼠瘫痪肢体功能的恢复。
Objective To study effects of neural stem cells（NSCs） transplantation on neurocyte apoptosis and bcl-2 expression of rat after spinal cord injury （SCI）. Methods 40 SD rats were divided into four groups randomly ： Normal group, SCI group, NSCs group and BrdU ＋ NSCs group. SCI model was induced with electrocircuit control spinal cord injuring device. NSCs in logarithmic phage were labeled with Brdu. The NSCs were transplanted into the injured site after SCI. The expression of Brdu and bcl-2 were detected by immunohistochemisty, and cell apoptosis of TUNEL were used to observe neural cell（ immunohistochemisty and immunofluorescence）. The improved Rivlin method （namely, slope test）was used to assess the motor function following spinal cord injury in rats. Results NSCs labeled with Brdu could be detected in the spinal cord after transplantation. TUNEL positive cells were significantly decreased at each time point in NSCs group and BrdU ＋ NSCs group than that of SCI group（P 〉0.01 ） ,the grey level of bcl-2 positive neurons at each time point in NSCs group and BrdU ＋ NSCs group were higher than those of SCI group （P 〈 0.01 ）, the expression peak of bcl-2 was postponed and reached the peak on 7thd. On 7thd, 14thd and 28thd after transplantation, inclined plane test of NSCs group and BrdU ＋ NSCs group were higher than those of SCI group（ P 〈 0.01 ）. There were no significant differences between NSCs group and BrdU ＋ NSCs group （ P 〉 0.05 ）. Conclusion The transplantation of NSCs can survive in the injured site of spinal cord, upregulate more obviously the expression of bcl-2 and inhibit neurocyte apoptosis after spinal cord injury, then promote more remarkably functional recovery of hind limb in rat after SC1.
出处
《中风与神经疾病杂志》
CAS
CSCD
北大核心
2012年第3期204-208,共5页
Journal of Apoplexy and Nervous Diseases
基金
山东省自然科学基金资助项目(ZR2010HM040)
山东省优秀中青年科学家科研奖励基金(博士基金)资助项目(BS2010YY006)
国家自然科学基金资助项目(81071779)
关键词
脊髓损伤
神经干细胞
细胞凋亡
Bcl-2
功能恢复
Spinal cord injury
Neural stem cells transplantation
Cell apoptosis
Bcl-2
Functional recovery
