中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2016, Vol. 11 ›› Issue (5): 846-853.doi: 10.4103/1673-5374.182764

• 原著:视神经损伤修复保护与再生 • 上一篇    下一篇

不同性别来源骨髓间充质干细胞对抗缺氧诱导的视神经节细胞凋亡的差异

  

  • 收稿日期:2015-12-22 出版日期:2016-05-20 发布日期:2016-05-20
  • 基金资助:

    国家自然科学基金（81100664）; 中山眼科中心眼科学国家重点实验室开放项目（303060202400306）; 武汉科技曙光计划（2014070404010222）; 武汉大学独立研究项目（2042014kf0259）

Gender difference in the neuroprotective effect of rat bone marrow mesenchymal cells against hypoxia-induced apoptosis of retinal ganglion cells

Jing Yuan1, Jian-xiong Yu2, *   

  1. 1 Eye Center, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China
    2 Department of Gastrointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China
  • Received:2015-12-22 Online:2016-05-20 Published:2016-05-20
  • Contact: Jing Yuan, M.D., Ph.D., xyj711@163.com.
  • Supported by:

    "This work was supported by grants from the National Natural Science Foundation of China, No. 81100664; the Open Project of the State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, China, No. 303060202400306; the Wuhan Science and Technology Dawn Project of China, No. 2014070404010222; and the Independent Research Project of Wuhan University of China, No. 2042014kf0259."

摘要：

骨髓间充质干细胞能减少视神经节细胞的死亡和有效地防止视力丧失。课题组以往研究中发现，雌性恒河猴骨髓间充质干细胞较雄性恒河猴可获得更强的神经分化潜能。因此，我们假设雌性骨髓间充质干细胞较雄性细胞可能具有更强的神经保护作用，实验首先经密度梯度离心法分离培养雌性和雄性成年大鼠骨髓间充质干细胞。经酶消化法获得新生大鼠视网膜组织来源的原代视神经节细胞。然后在缺氧条件下将视神经节细胞与骨髓间充质干细胞在transwell系统中进行共培养。采用流式细胞仪测定和caspase-3活性下检测细胞的凋亡，后发现视神经节细胞凋亡率和caspase-3活性在缺氧24 h较常氧条件明显增高；当视神经节细胞与雌性和雄性鼠来源的骨髓间充质干细胞在缺氧条件下共培养时，其细胞凋亡率和caspase-3活性均较单独培养显著下降，且以与雌性鼠来源的干细胞共培养作用更明显。结果证实了骨髓间充质干细胞可抑制缺氧诱导视神经节细胞凋亡，且雌性来源的干细胞具有更强神经保护作用的假设。 

orcid: 0000-0001-6757-0217(Jing Yuan)

关键词: 神经再生, 视神经损伤, 骨髓间充质干细胞, 视神经节细胞, 神经保护, 缺氧损伤, 性别差异, Transwell 系统, 共培养, 细胞凋亡, 流式细胞仪检测, Caspase-3

Abstract:

"Bone marrow mesenchymal stem cells can reduce retinal ganglion cell death and effectively prevent vision loss. Previously, we found that during differentiation, female rhesus monkey bone marrow mesenchymal stem cells acquire a higher neurogenic potential compared with male rhesus monkey bone marrow mesenchymal stem cells. This suggests that female bone marrow mesenchymal stem cells have a stronger neuroprotective effect than male bone marrow mesenchymal stem cells. Here, we first isolated and cultured bone marrow mesenchymal stem cells from female and male rats by density gradient centrifugation. Retinal tissue from newborn rats was prepared by enzymatic digestion to obtain primary retinal ganglion cells. Using the transwell system, retinal ganglion cells were co-cultured with bone marrow mesenchymal stem cells under hypoxia. Cell apoptosis was detected by flow cytometry and caspase-3 activity assay. We found a marked increase in apoptotic rate and caspase-3 activity of retinal ganglion cells after 24 hours of hypoxia compared with normoxia. Moreover, apoptotic rate and caspase-3 activity of retinal ganglion cells significantly decreased with both female and male bone marrow mesenchymal stem cell co-culture under hypoxia compared with culture alone, with more significant effects from female bone marrow mesenchymal stem cells. Our results indicate that bone marrow mesenchymal stem cells exert a neuroprotective effect against hypoxia-induced apoptosis of retinal ganglion cells, and also that female cells have greater neuroprotective ability compared with male cells."

Key words: nerve regeneration, optic nerve injury, bone marrow mesenchymal stem cells, retinal ganglion cells, neuroprotection, hypoxic injury, gender difference, transwell system, co-culture, cell apoptosis, flow cytometry, caspase-3, neural regeneration