摘要
目的探讨大鼠脑出血微创后行神经干细胞移植对其神经功能恢复的影响。方法（1）从新生24h内的Wistar乳鼠海马组织中分离出神经干细胞，利用无血清培养技术，在体外进行传代培养、标记和鉴定。（2）采用未肝素化的心脏自体血制作大鼠尾状核脑出血模型。将模型分为正常对照组、脑出血自然恢复组、脑出血微创组、脑出血移植组、脑出血微创＋移植组。（3）模型成功后24h行立体定向脑内血肿吸引清除术，出血后48h将体外扩增、纯化、并经Brdu标记的神经干细胞立体定向移植于血肿残腔内，不同时间点观察评估各组大鼠神经行为学改善情况。（4）通过免疫组化及荧光双标检测移植入血肿残腔的神经干细胞在脑出血周边组织中的增殖、分化情况。结果（1）大鼠脑出血后神经功能评分，脑出血自然恢复组在术后21d与3d比较明显减低（P〈0．05），脑出血微创组、脑出血移植组及脑出血微创＋移植组在术后14d、21d与3d比较明显减低（P〈0．05），脑出血微创组及脑出血移植组仅在术后21d时的评分明显低于脑出血自然恢复组（P〈0．05），而脑出血微创＋移植组在术后14d时的评分即较脑出血自然恢复组明显减低（P〈0．05），其术后21d的评分也明显低于脑出血微创组和移植组（P〈0．05），脑出血微创组与移植组各时间点的评分相比，差异无统计学意义（P〉0．05）。（2）大鼠脑出血周边组织中BrdU阳性细胞数在出血后7—14d明显增多，14d时达到高峰，21d时明显减少。除3d外，在7d、14d和21d的3个时间点，脑出血微创＋移植组BrdU阳性及NSE／BrdU双标阳性细胞数均多于脑出血移植组，差异具有统计学意义（P〈0．05）。结论从新生Wistar乳鼠海马组织中分离培养的是神经干细胞，并在体外能够长期培养和传代。大鼠脑出血微创后行神经干细胞移植能明显促进其神经功能的恢复。微创对神经干细胞在脑出血周边组织中的增殖、分化有促进作用。
Objective To investigate the effect of nerve function in rats with cerebral hemorrhage after minimal inva- sion across neural stem cell transplantation Methods （ 1 ） Hippocampus neural stem cells was isolated from newborn Wist- ar rat, neural stem cells was cultured, marked and identificatied in vitro by using serum-free culture technology. （ 2 ） the model of cerebral hemorrhage of rat was produced by using the non heparinized autologous blood from the heart of the cau- date nucleus, the models were divided into the following 5 groups：control group, spontaneous recovery group, minimally in- vasive group, transplant group and minimally invasive surgery after the transplant group. （ 3 ） stereotactic intracerebral hema- toma attract dissection was made after 24h, Amplificatied and purificatied neural stem cells after BrdU labeled were trans- plantted into stereotactic residual hematoma cavity after 48h, assessment of the neurobehavior was observed at different time points. （4）the proliferation and differentiation of neural stem cells after transplantation into hematoma cavity was detected by immunohistochemistry and double-labeled fluorescence. Results The score of neurological functions was made. the score of 21d was significantly lower than the 3d＇ s in spontaneous recovery group（ P 〈0.05 ） ,the scores of 14d and 21d of minimally invasive group, transplant group and minimally invasive surgery after the transplant group were significantly lower than the 3 d＇ s in same group （ P 〈 0.05 ）. the scores of the 21d of minimally invasive group and transplant group were sig- nificantly lower than the spontaneous recovery group＇s in 21d（P 〈 0.05 ） ,while the score of the 14d of minimally invasive surgery after the transplant group was lower than that of 14d of spontaneous recovery group（ P 〈 0.05 ） , and the score of the 21 d of minimally invasive surgery after the transplant group was lower than that of 21d of minimally invasive group eand transplant group（ P 〈 0.05 ） ,there were no significant difference between the scores of minimally invasive group and trans- plant group at different time（ P 〉 0.05 ）. 2 ,the number of BrdU ＋ cells in rat brain tissue surrounding hematoma increased significantly after 7d, and increase,reach the peak in 14d,and then decrease in 21d. the number of BrdU ＋ and NSE/BrdU double-labeled-positive cells of transplantation after minimally invasive group were more than that of transplant group in 7d, 14d and 21d in addition to 3d（ P 〈 0.05 ）. Conclusions neural stem cells can be isolated from the hippocampus of new- born Wistar rat, can be cultured in vitro for a long-term, transplantation of neural stem cells after undergo minimally inva- sion can promote the recovery of neurological function after cerebral hemorrhage. Minimally invasion promote the prolifera- tion and differentiation of neural stem cell in cerebral hemorrhage
出处
《中风与神经疾病杂志》
CAS
CSCD
北大核心
2013年第11期1012-1015,共4页
Journal of Apoplexy and Nervous Diseases
基金
黑龙江省教育厅科学研究项目(12511561)
关键词
微创
脑出血
神经干细胞
移植
Minimally invasion
Cerebral Hemorrhage
Neural stem cells
Transplantation
