中国神经再生研究(英文版) ›› 2015, Vol. 10 ›› Issue (12): 2011-2017.doi: 10.4103/1673-5374.172320

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

体外构建血脑屏障模型:用于研究神经保护药物的作用机制

  

  • 收稿日期:2015-07-27 出版日期:2015-12-30 发布日期:2015-12-30
  • 基金资助:

    中国国家自然科学基金项目(81374005, 30973979),十二五计划项目(2012BAI26B03)

In vitro model of the blood-brain barrier established by co-culture of primary cerebral microvascular endothelial and astrocyte cells

Yan Wang, Ning Wang*, Biao Cai, Guang-yun Wang, Jing Li, Xing-xing Piao   

  1. Key Laboratory of Xin’an Medicine, Ministry of Education; Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine, Anhui Academy of Chinese; College of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui Province, China
  • Received:2015-07-27 Online:2015-12-30 Published:2015-12-30
  • Contact: Ning Wang, Ph.D., wnhefei@163.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81374005, 30973979; a grant from the National Science and Technology Support Program during the Twelfth “Five-Year” Plan Period of China, No. 2012BAI26B03.

摘要:

由于治疗神经系统疾病的药物需要渗透血脑屏障,因此建立体外血脑屏障模型是探索药物渗透的关键。但目前建立血脑屏障模型的方法尚不完善。我们以0.4μm孔径微孔膜的Transwell小室作为支持,将来源于新生大鼠脑组织中的脑微血管内皮细胞和星型胶质细胞分别种植于其微孔膜的内外侧,建立体外血脑屏障模型。跨上皮电阻、泄漏试验和特异性酶检测结果显示,构建的血脑屏障模型具有体内血脑屏障相同的低渗透特征,实验者可使用这种模型研究神经保护药物的作用机制。

关键词: 神经再生, 血脑屏障, 微血管内皮细胞, 星形胶质细胞, 渗透性, 共培养, Transwell小室

Abstract:

Drugs for the treatment and prevention of nervous system diseases must permeate the blood-brain barrier to take effect. In vitro models of the blood-brain barrier are therefore important in the investigation of drug permeation mechanisms. However, to date, no unified method has been described for establishing a blood-brain barrier model. Here, we modified an in vitro model of the blood-brain barrier by seeding brain microvascular endothelial cells and astrocytes from newborn rats on a polyester Transwell cell culture membrane with 0.4-μm pores, and conducted transepithelial electrical resistance measurements, leakage tests and assays for specific blood-brain barrier enzymes. We show that the permeability of our model is as low as that of the blood-brain barrier in vivo. Our model will be a valuable tool in the study of the mechanisms of action of neuroprotective drugs.

Key words: nerve regeneration, blood-brain barrier, astrocytes, brain microvascular endothelial cells, permeability, co-culture, Transwell chamber, neural regeneration