兰州大学机构库 >学院待认领
骨髓基质干细胞移植联合应用G-CSF对大鼠脊髓损伤修复的影响
Alternative TitleEffects of Combination of Bone Marrow Stromal Stem Cells Transplantion with Granulocyte-Colony Stimulating Factor on Repair of Rat Spinal Cord Injury
李晓飞
Thesis Advisor文益民
2011-05-18
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword脊髓损伤 骨髓基质干细胞 粒细胞集落刺激因子 移植
Abstract目的: 探讨骨髓基质干细胞(Bone marrow stromal stem cells,BMSCs)移植联合应用粒细胞集落刺激因子(granulocyte colony stimulating factor,G-CSF)对大鼠急性脊髓损伤(spinal cord injury,SCI)的治疗修复作用。 方法: 取8周龄大小的4只健康Wistar大鼠骨髓,分离、培养BMSCs,在移植前24小时取第三代BMSCs行Hoechst33342标记,将标记细胞调至1×105/ul制备成BMSCs悬液备用。48只Wistar大鼠采用自制改良的Allen's装置在T11水平制成大鼠急性脊髓损伤模型,根据实验设计要求将动物模型随机分成4组(n=12):A组为BMSCs移植联用G-CSF治疗组,B、C组为单纯BMSCs移植和G-CSF治疗组,D组为损伤对照组。术后1、2、3、4周采用Basso-Beattie-Bresnahan(BBB)评分评价大鼠后肢神经功能恢复情况;术后4周取材用荧光显微镜在损伤区及周缘观察BMSCs存活、分化情况,并行HE染色作大体组织学观察;免疫荧光染色检测神经元特异性烯醇化酶(neuron-specific enolase,NSE)、神经胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和神经丝蛋白200(neurofilament 200,NF-200)的表达变化。 结果:术后1~4周,A组大鼠BBB评分呈持续升高趋势,A组评分均明显高于其它三组,D组最低,差异均有统计学意义(P<0.01);B组术后3、4周高于C组(P<0.01);B、C组与D组比较差异有统计学意义(P< 0.01)。HE染色显示,BMSCs联合应用G-CSF对脊髓损伤的修复作用最好,D组恢复最差,B、C组介于A、D组之间;免疫荧光染色示,术后4周在SCI损伤区及周缘可见大量Hoechst 33342标记的BMSCs聚集,灰质分布较白质多,BMSCs从移植部位向周围损伤组织迁移,A组存活细胞数高于B组;A组在脊髓损伤区及周缘NSE、NF200阳性细胞均较B组多,C组未见明显的NSE、NF200阳性细胞,但在损伤周缘有大量的GFAP阳性细胞,并向脊髓损伤空腔内延伸;D组损伤区看见大量结构杂乱的GFAP阳性细胞,瘢痕组织形成明显,损伤区未见明显的脊髓再生现象及NSE、NF-200阳性细胞。
Other AbstractObjective To investigate effects of transplantation of bone marrow stromal stem cells (BMSCs) combined with granulocyte colony stimulating factor(G-CSF)on repair of the acute spinal cord injury (SCI) in adult rat. Methods BMSCs were separated and cultured from four Wistar rats aged 8 weeks. 24h before transplanting, BMSCs of the third generation were labeled by Hoechst 33342, then adjusted the labeled BMSCs to 1×105, prepared monoplast suspension for use. 48 adult wistar rats were made the acute spinal cord injury (SCI) models at T11 level according to the improved Allen’s method and the requirement of experiment, the qualified animals were randomly divided into four groups(n=12):BMSCs and G-CSF treatment group (group A), BMSCs transplantation group (group B),G-CSF treatment group (group C), single SCI group (group D).After 1, 2, 3 and 4 weeks of surgery, the functional recovery of the hind limbs was evaluated by the Basso-Beattie-Bresnahan (BBB) locomotor rating score. after 4 weeks of surgery, Specimens were procured at 4 weeks after surgery for investigating the survival and differentiation of BMSCs by fluorescence microscope in the damaged area and border area. In addition, gross anatomy and histology were observed by HE staining.Immunofluorescence measurement were used to detected expression of neuron-specific enolase(NSE),glial fibrillary acidic protein(GFAP)and neurofilament 200(NF-200)proteins. Results 1-4 weeks after surgery , the values of BBB in group A continually increased,the BBB scores in group A were significantly higher than other groups, Group D was the lowest, the differences were statistically significant (P <0.01);B Group higher than Group C at 3,4 weeks (P <0.01), Group D performed significant difference with group B and group C(P< 0.01).HE staining revealed that BMSCs combined with G-CSF on repair of spinal cord injury were best, Group D were the worst, Group B、C ranged between A and D groups. 4 weeks after surgery, Immunofluorescent staining results showed there were a great quantity of BMSCs gathered which were labeled by Hoechst 33342 in the SCI damaged area and border area in gray matter more than white matter, migrated from the transplantation spot to the injured tissue,The number of survival cell in group A was more than group B.The NSE,GFAP and NF 200-positive cells around the SCI in the group A were higher than group B. The NSE, NF 200-positive cells were no obvious , but GFAP-positive cells ...
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223384
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
李晓飞. 骨髓基质干细胞移植联合应用G-CSF对大鼠脊髓损伤修复的影响[D]. 兰州. 兰州大学,2011.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Altmetrics Score
Google Scholar
Similar articles in Google Scholar
[李晓飞]'s Articles
Baidu academic
Similar articles in Baidu academic
[李晓飞]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[李晓飞]'s Articles
Terms of Use
No data!
Social Bookmark/Share
No comment.
Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.