中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (45): 8416-8420.doi: 10.3969/j.issn.1673-8225.2010.45.014

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

沉默RhoA基因对骨髓间充质干细胞静脉移植治疗脑梗死大鼠的作用

周国庆，金  怡，张  鹏   

  1. 中国石油集团东方物理勘探有限责任公司职工中心医院，河北省保定市  072550
  • 出版日期:2010-11-05 发布日期:2010-11-05
  • 作者简介:周国庆，男，1975年生，河北省徐水县人，汉族，1999年河北医科大学毕业，主治医师，主要研究脑血管病的防治与治疗。 phoenix1a1@163.com

Treatment of cerebral infarction in rats by transplanting bone marrow mesenchymal stem cells with RhoA gene silencing via the vein

Zhou Guo-qing, Jin Yi, Zhang Peng   

  1. Oil Geophysical Prospecting Bureau of Trade Unions Central Hospital， Baoding  072550， Hebei Province， China
  • Online:2010-11-05 Published:2010-11-05
  • About author:Zhou Guo-qing， Attending physician， Oil Geophysical Prospecting Bureau of Trade Unions Central Hospital， Baoding 072550， Hebei Province， China phoenix1a1@163.com

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摘要：

背景：研究证实，RhoA基因在神经继发损伤中发挥着重要的作用，通过基因沉默降低其表达，可以有效阻断继发性神经损伤。RhoA蛋白介导的神经突生长抑制作用是影响骨髓间充质干细胞移植后修复效果的重要原因。
目的：验证是否可以通过沉默RhoA基因的方法，改良骨髓间充质干细胞静脉移植对大鼠脑梗死的治疗效果。
方法：体外培养骨髓间充质干细胞，经小分子干扰RNA转染以沉默RhoA基因表达，用Western blot法检测神经干细胞在转染前后RhoA基因的表达。建立大脑中动脉闭塞模型，分为3组，通过尾静脉分别注入RhoA基因沉默的骨髓间充质干细胞悬液、骨髓间充质干细胞悬液及不含干细胞的培养液(对照组)。注入移植液后24 h，3 d，1，2 周行Bederson评分检测神经功能的损伤情况。造模2周后处死大鼠取材，行免疫组织化学和苏木精-伊红染色观察大脑梗死处组织学变化。
结果与结论：经分子干扰RNA转染后24 h的骨髓间充质干细胞RhoA基因表达较转染前显著降低(P=0.002)。沉默RhoA基因后的骨髓间充质干细胞移植治疗，无论从组织学上还是功能学上疗效都有明显的提高。提示用RNA干扰的方法使骨髓间充质干细胞的RhoA基因沉默，再通过尾静脉注射的方法移植到大鼠颅脑损伤区，移植的骨髓间充质干细胞可以更好地在损伤部位存活，增殖分化与迁移，促进脑梗死后大鼠神经功能的恢复。

关键词: 脑梗死, 骨髓间充质干细胞, 干细胞移植, RhoA, RNA干扰

Abstract:

BACKGROUND: Previous studies have confirmed that RhoA gene plays an important role in nerve damages. To decrease its expression by gene silencing can effectively block secondary nerve damages. RhoA protein mediated neurite growth inhibitory effect is an important reason for affecting repair effect following bone marrow mesenchymal stem cells (BMSCs) transplantation.
OBJECTIVE: To determine whether the RhoA gene silencing in BMSCs can enhance curative effects of transplantation by vein of BMSCs on cerebral infarction in rats.
METHODS: BMSCs were cultured by the suspension culture in vitro, and were transfected by siRNA to knock down the expression of RhoA gene. Western blot assay was used to assess the knockdown efficiency. The models of cerebral infarction were made by middle cerebral artery occlusion and randomly divided into three groups. RhoA gene silencing of BMSCs suspension, BMSCs suspension and medium without stem cells (control group) were respectively injected via caudal vein. The neurological defect scores were determined at 24 hours, 3 days and 1, 2 weeks after injection. The rats were sacrificed at 2 weeks following model induction. Histological changes in infarct regions were observed using immunohistochemistry and hematoxylin-eosin staining.
RESULTS AND CONCLUSION: The expression of RhoA was markedly knocked down by transfection of siRNA at 24 hours after the transfection (P=0.002). Therapeutic effects of BMSCs transplantation following RhoA gene silencing were significantly increased in histology and function. These suggest that RhoA gene in BMSCs can be silenced by RNA interference. BMSCs of RhoA gene silencing transplanted by caudal vein into the cerebral infarction tissues can survive, proliferation, differentiate and migrate in damage sites, and significantly improve the neurological function in rats with cerebral infarction.

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