中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (4): 597-604.doi: 10.4103/1673-5374.247464

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

人骨髓间充质干细胞对脑缺血神经保护作用可由抗细胞凋亡机制介导

  

  • 出版日期:2019-04-15 发布日期:2019-04-15
  • 基金资助:
    NIH R01NS071956，NIH R01 NS090962，NIH R21NS089851，NIH R21 NS094087和VA Merit Review I01 BX001407资助

Neuroprotective effects of human bone marrow mesenchymal stem cells against cerebral ischemia are mediated in part by an anti-apoptotic mechanism

Yuyang Zhang 1, 2 , Seongjin Yu 1 , Julian P. Tuazon 1 , Jea-Young Lee 1 , Sydney Corey 1 , Lauren Kvederis 1 , Chase Kingsbury 1 , Yuji Kaneko 1 , Cesar V. Borlongan 1   

  1. 1 Department of Neurosurgery and Brain Repair, Center of Excellence for Aging and Brain Repair, University of South Florida, Tampa, FL, USA
    2 Department of Pharmacology / School of Life Science and Biopharmaceutical Science, Shenyang Pharmaceutical University, Shenyang, Liaoning Province, China
  • Online:2019-04-15 Published:2019-04-15
  • Contact: Cesar V. Borlongan, PhD, cborlong@health.usf.edu.
  • Supported by:

    Cesar V. Borlongan was funded by NIH R01NS071956, NIH R01 NS090962, NIH R21NS089851, NIH R21 NS094087, and VA Merit Review I01 BX001407.

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摘要：

人骨髓间充质干细胞（hMSCs）移植已被证实对脑卒中有治疗作用，但其确切机制尚不清楚。为了解人骨髓间充质干细胞对脑缺血的神经保护作用是否与抗细胞凋亡机制有关。实验将大鼠神经元和星形胶质细胞的原代混合培养物暴露于氧糖剥夺条件下，并在标准培养基和含氧量正常的条件下进行2h的“再灌注”，然后立即进行人骨髓间充质干细胞和/或Bcl-2抗体处理。（1）通过MTT和台盼蓝排除法测定原代大鼠神经元和星形胶质细胞的细胞活力；（2）通过免疫细胞化学表征人骨髓间充质干细胞存活和分化；（3）通过酶联免疫吸附法测定上清液中Bcl-2的浓度；（4）结果可见：培养的人骨髓间充质干细胞表达胚胎样干细胞表型标志物CXCR4，Oct4，SSEA4和Nanog，以及未成熟的神经表型标志物巢蛋白；人骨髓间充质干细胞可使原代大鼠神经元和星形胶质细胞免受氧糖剥夺侵害，这种作用被Bcl-2抗体拮抗；但在上清液中的Bcl-2水平在暴露于氧糖剥夺的人骨髓间充质干细胞和未处理细胞之间无差异，可以解释人骨髓间充质干细胞对脑缺血神经保护的作用部分由抗细胞凋亡机制介导。

orcid: 0000-0002-2966-9782(Cesar V. Borlongan)

关键词: 人骨髓间充质干细胞, 脑卒中, 脑缺血, 凋亡, Bcl-2抗体, 氧糖剥夺, 神经保护

Abstract:

Transplantation of human bone marrow mesenchymal stem cells (hMSCs) stands as a potent stroke therapy, but its exact mechanism remains unknown. This study investigated the anti-apoptotic mechanisms by which hMSCs exert neuroprotective effects on cerebral ischemia. Primary mixed cultures of rat neurons and astrocytes were cultured and exposed to oxygen-glucose deprivation. A two-hour period of “reperfusion” in standard medium and normoxic conditions was allowed and immediately followed by hMSCs and/or Bcl-2 antibody treatment. Cell viability of primary rat neurons and astrocytes was determined by 3-(4,5-dimethylthianol-2-yl)-2,5 diphenyl tetrazolium bromide and trypan blue exclusion methods. hMSC survival and differentiation were characterized by immunocytochemistry, while the concentration of Bcl-2 in the supernatant was measured by enzyme-linked immunosorbent assay to reveal the secretory anti-apoptotic function of hMSCs. Cultured hMSCs expressed embryonic-like stem cell phenotypic markers CXCR4, Oct4, SSEA4, and Nanog, as well as immature neural phenotypic marker Nestin. Primary rat neurons and astrocytes were protected from oxygen-glucose deprivation by hMSCs, which was antagonized by the Bcl-2 antibody. However, Bcl-2 levels in the supernatants did not differ between hMSC- and non-treated cells exposed to oxygen-glucose deprivation. Neuroprotective effects of hMSCs against cerebral ischemia were partially mediated by the anti-apoptotic mechanisms. However, further studies are warranted to fully elucidate this pathway.

Key words: apoptosis, Bcl-2 antibody, human mesenchymal stem cells, ischemia, neuroprotection, oxygen glucose deprivation