中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (40): 7462-7469.doi: 10.3969/j.issn.1673-8225.2010.40.012

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

Rolipram联合少突胶质前体细胞移植治疗大鼠脊髓损伤

祁  全1,毕郑钢1,杨卫良1,李学钊1,杨  鑫1,张  磊2,潘尚哈3   

  1. 哈尔滨医科大学附属第一医院,1骨科,3中心实验室,黑龙江省哈尔滨市 150001;2哈尔滨医科大学病理教研室,黑龙江省哈尔滨市150001
  • 出版日期:2010-10-01 发布日期:2010-10-01
  • 通讯作者: 毕郑钢,硕士,教授,哈尔滨医科大学附属第一医院骨科,黑龙江省哈尔滨市 150001 bizhenggang@54dr.com
  • 作者简介:祁全☆,男,1972年生,黑龙江省哈尔滨市人,汉族,2006年哈尔滨医科大学毕业,博士,副主任医师,主要从事脊柱脊髓损伤方面的研究。 qiquan999@sina.com
  • 基金资助:

    黑龙江省青年基金资助项目(QC06C073);黑龙江省卫生厅基金资助项目(2006-078);黑龙江省教育厅基金资助项目(10551170);黑龙江省博士后资助项目;哈医大一院科研基金资助项目(2007037)。

Rolipram combined with oligodendrocyte precursor cell transplantation for treatment of spinal cord injury in rats

Qi Quan1, Bi Zheng-gang1, Yang Wei-liang1, Li Xue-zhao1, Yang Xin1, Zhang Lei2, Pan Shang-ha3   

  1. 1 Department of Orthopaedics, 3 Central Laboratory, First Affiliated Hospital, Harbin Medical University, Harbin  150001, Heilongjiang Province, China; 2 Department of  Pathology, Harbin Medical University, Harbin  150001, Heilongjiang Province, China
  • Online:2010-10-01 Published:2010-10-01
  • Contact: Bi Zheng-gang, Master, Professor, Department of Orthopaedics, First Affiliated Hospital, Harbin Medical University, Harbin 150001, Heilongjiang Province, China bizhenggang@54dr.com
  • About author:the Youth Foundation of Heilongjiang Province, No. QC06C073*; the Grant of Health Department of
  • Supported by:

    Heilongjiang Province, No. 2006-078*; the Grant of Education Department of Heilongjiang Province, No. 10551170*; the Postdoctoral Foundation of Heilongjiang Province*; the Scientific Research Foundation of First Hospital of Harbin Medical University, No. 2007037*

摘要:

背景:脊髓损伤轴突难以再生涉及到多方面原因,综合运用多种治疗方法应该更有效。
目的:应用Rolipram联合少突胶质前体细胞移植治疗大鼠脊髓损伤,观察轴突再生和再髓鞘化的情况。
方法:Wistar大鼠利用ALLEN法打击形成脊髓损伤模型。Rolipram治疗组和联合治疗组背部皮下埋置ALZET渗透性微泵内装Rolipram 200 μL,每小时释放0.5 μL,2 mg/(kg•d),可持续14 d,1周后局部注入0.012 5 μmol 的双丁酸环腺苷酸;细胞移植组和联合治疗组移植少突胶质前体细胞,脊髓损伤组注入同等量的生理盐水,术后每周测定BBB运动评分,4周取材,冰冻切片,苏木精-伊红染色,免疫荧光染色。
结果与结论:伤后4周Rolipram治疗组和联合治疗组BBB运动评分高于脊髓损伤组和细胞移植组;免疫荧光显示Rolipram治疗组和联合治疗组损伤局部NF200表达旺盛;细胞移植组和联合治疗组少突胶质前体细胞存活并表达髓鞘碱性蛋白,后者存活数量更多,并且有髓纤维增多。说明应用Rolipram提高环磷腺苷水平促进了大鼠脊髓损伤功能的恢复,联合少突胶质前体细胞移植可产生协同促进作用。

关键词: 脊髓损伤, 环磷腺苷, 少突胶质前体细胞, 细胞移植, 协同作用

Abstract:

BACKGROUND: There are many reasons for the difficult regeneration of the injured spinal cord axon, and a comprehensive application of several methods will be more effective.
OBJECTIVE: To observe axonal regeneration and remyelinization using Rolipram combined with oligodendrocyte precursor cells (OPCs) transplantation in rats after spinal cord injury (SCI).
METHODS: Wistar rat models of SCI were established using ALLEN method. Alzet osmotic pumps contained Rolipram 200 μL were embed subcutaneouly in Rolipram treatment and combination groups, release 0.5 μL per hour, 2 mg/(kg•d) for 14 days. After one week, 0.012 5 μmol dibutyryl cyclic-cyclic adenosine monophosphate was infused in the injured spinal cord. OPCs were infused to cell transplantation and combination groups. An equal volume of saline was injected in the SCI group. Basso Beattie and Bresnahan score was assessed weekly postoperation until 4 weeks when all rats were killed and spinal cords were made into frozen sections. Hematoxylin-eosin staining and immunofluorescence method were performed.
RESULTS AND CONCLUSION: At 4 weeks postoperation, Basso Beattie and Bresnahan score was greater in the Rolipram treatment and combination groups compared with SCI and cell transplantation groups. Immunofluorescence method showed: the expression of NF200 was strong in Rolipram treatment and combination groups. OPCs were alive and positive for myelin basic protein, which was predominant and myelinated nerve fiber increased in latter. These suggest that Rolipram elevated cyclic adenosine monophosphate levels, promoted functional recovery after SCI, especially combined with OPC transplantation.

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