摘要
目的:评估脂肪间充质干细胞(adipose derived mesenchymal stem cells,ADSCs)移植对脊髓损伤(spinal cord injury,SCI)小鼠血小板反应蛋白4(thrombospondin-4,Thbs-4)表达水平的影响,及ADSCs修复SCI的作用。方法:制备小鼠T10 SCI模型,随机分为假手术组、模型组、ADSCs移植组,每组30只。取同种系转绿色荧光基因小鼠皮下脂肪组织,分离培养ADSCs。损伤后即刻移植106个细胞至ADSCs移植组小鼠脊髓内,模型组注射同等剂量PBS。术前及术后每周进行运动功能BBB评分;采用免疫荧光法检测移植细胞存活、炎症反应情况;采用Western Blot法检测移植术后第7 d各组动物脊髓组织Thbs-4蛋白表达水平;采用ELISA法检测术后第3、14 d各组动物脊髓组织TNF-α浓度。结果:ADSCs移植后可在宿主体内存活至少2周。移植术后第7 d,Thbs-4在假手术组表达水平最低,模型组次之,在ADSCs移植组表达水平最高(P<0.05);ADSCs移植下调SCI后TNF-α浓度;移植术后第14 d,ADSCs移植组ED-1+炎症细胞数量显著少于模型组(P<0.05);运动功能评分结果显示从第2周起,ADSCs移植组小鼠显著优于模型组(P<0.05)。结论:ADSCs移植促进SCI小鼠运动功能恢复,其机制与提升Thbs-4蛋白表达水平,减轻受损脊髓组织炎症反应增生有关。
Objective： To investigate the effects of transplantation of adipose derived mesenchymal stem cells on the expression of thrombospondin-4（ Thbs-4） and the recovery of locomotor function after acute spinal cord injury（ SCI） in mice. Methods： The spinal cord was crush-injured at the T10 level in mice. SCI mice were randomly divided into 2 experimental groups（ thirty mice each group） ： a control group receiving PBS（ control group）; a group receiving ADSCs（ ADSCs group）,a sham group was also included（ sham group,n = 30）. 106 ADSCs derived from GFP-transgenic mice of the same strain were transplanted into the injured spinal cord immediately after injury,the control group received the same dosage PBS. Locomotor recovery was assessed with BBB scoring pre-operation and post-operation weekly; the sur-vival of grafted cells and the inflammation reaction were detected by immunofluorescence method. The expression of Thbs-4 was detected by Western Blot at 7 days post-injury; The concentration of TNF-α on the extracts from the spinal cord were examined by ELISA at 3 days and 14 days post-injury. Results： Transplanted ADSCs survived in the host at least 14 days. 7 days post transplantation,compared with the sham group,the expression of Thbs-4 was significantly increased in the control group（ P〈0. 05）,ADSCs transplantation further enhanced the expression of Thbs-4（ P〈0. 05）. Compared to control group,transplants with ADSCs significantly decreased the number of ED-1 ＋ cells,downregulated the concentration of TNF-α in the injured spinal cord（ P〈0. 05）. Compared with control group,the BBB score in ADSCs group showed marked improvement between the 2 th week and the 4 th week after injury（ P〈0. 05）.Conclusion： ADSCs transplantation improves the locomotors recovery of SCI mice,the underlying mechanism involves the up-regulation the expression of Thbs-4 and down-regulation of inflammation.
作者
周治来
徐华丽
姚舜
黄子祥
黄炫武
黄帅
许立惟
张辉
Zhou Zhilai;Xu Huali;Yao Shun;Huangzixiang;Huang Xuanwu;Huang Shuai;Xu Liwei;Zhang Hui(Department of Spine Surgery, Guangdong Second Provincial General Hospital, Guangzhou 510317;Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282;Graduate School, Southern Medical University, Guangzhou 510515, China)
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2018年第3期371-376,共6页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金-青年科学基金(81400996)
广东省自然科学基金(2016A030313627)
