天津医药

• 实验研究 •    

骨髓间充质干细胞NgR基因沉默静脉移植治疗大鼠脑损伤

王东   

  1. 天津第四中心医院
  • 收稿日期:2009-12-29 修回日期:2010-06-13 出版日期:2010-09-15 发布日期:2010-09-15
  • 通讯作者: 王东

Treatment of traumatic brain injury in rats by transplanting of BMSCs with NgR gene Silencing by vein

wang dong   

  • Received:2009-12-29 Revised:2010-06-13 Published:2010-09-15 Online:2010-09-15
  • Contact: wang dong

PDF (PC)

1850

摘要： 【摘 要】 目的：检测是否可以通过沉默NgR基因的方法提高骨髓间充质干细胞静脉移植对大鼠脑损伤的治疗效果。方法：体外培养骨髓间充质干细胞,经小分子干扰RNA(small interfering RNA, siRNA)转染以沉默NgR基因表达,用Western blot检测神经干细胞在转染前后NgR基因蛋白的表达采用液压颅脑损伤仪，给予2．5—3．0 atm液压冲击力，制成重型液压颅脑损伤模型，分为三组, A 组为NgR基因沉默的骨髓间充质干细胞组B 组为等量的骨髓间充质干细胞悬液组。C组为注射等量的不含干细胞的培养液组。静脉移植后24 h ，3天及伤后1、2 周行Bederson评分检测神经功能的损伤情况。2 周后处死行免疫组化和HE染色。结果：转染siRNA后，与对照组相比，实验组NgR基因蛋白表达量明显降低，移植后1周，A 组，B 组大鼠神经学缺损评分明，A 组低于B 组，B 组低于C组（P<0.05）；2周后其脑组织组化和HE切片中的神经元数量和NF阳性纤维A 组多于B 组，B 组多于C组（P<0.05）；结论：骨髓间充质干细胞NgR基因沉默静脉移植治疗大鼠脑损伤可明显改大鼠的神经学功能。

关键词: 脑损伤, 骨髓间充质干细胞, NgR, RNA干扰

Abstract: 【Abstract】Objective To determine whether the NgR 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 NgR. Western blot and were used to assess the knockdown efficiency. The models of cerebral infarction were made by Middle cerebral artery occlusion (MCAQ) and randomly divided into three groups, A group for the NgR gene silencing of BMSCs .B Group for the equivalent group of BMSCs suspension. C group was injected the same amount of non-stem cell of the culture medium. The neurological defect scores were determined after 24 h, 3 days and 1, 2 weeks after MCAQ. The immunohistochemistry and pathology were checked after 2 weeks . 【Result】Western blot approved that the expression of NgR was knocked down by transfection of siRNA at 24 hours after the transfection. The neurological defect scores were significantly lower in A group than those in the B and C group 1 weeks after MCAQ (P<0.05). The number of neurons in A group was higher than that in the B and C group(P<0.05) .Conclusion: The BMSCs of NgR gene silencing transplanted by vein into the cerebral infarction tissues can significantly improve the neurological function in the rats with TBI