中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (15): 2769-2772.doi: 10.3969/j.issn.1673-8225.2011.15.026

• 组织构建与生物活性因子 tissue construction and bioactive factors • 上一篇    下一篇

脑源性神经营养因子与心脏微血管内皮细胞的管状形成

刘少坤1,2,3，曹  亮1,2,3，张  靓1,2,3，沈晓涛1,2,3，赵宝寅1,2,3，齐绪峰1,2,3，秦俊文1,4，蔡冬青1,2,3   

  1. 暨南大学，1再生医学教育部重点实验室，2再生医学香港中文大学-暨南大学联合实验室，3科技部及广东省科技厅国际科技合作基地，4生殖免疫研究所，广东省广州市  510632
  • 收稿日期:2010-12-23 修回日期:2011-03-05 出版日期:2011-04-09 发布日期:2013-11-06
  • 通讯作者: 蔡冬青，教授，博士生导师，暨南大学，再生医学教育部重点实验室，再生医学香港中文大学-暨南大学联合实验室，科技部及广东省科技厅国际科技合作基地，广东省广州市 510632 tdongbme@jnu. edu.cn
  • 作者简介:刘少坤★，男，1983年生，山东省泰安市人，汉族，暨南大学在读硕士，主要从事心脏内皮细胞增殖、迁移、血管新生与管状形成方面的研究。 sakuelee@gmail.com
  • 基金资助:

    863项目(2007AA02Z105)；国家自然科学基金(30770886，30570369，30340038，30973158)；广东省自然科学基金重点项目(04105826)；广东省科技攻关项目(2004B30601007)；广东省国际合作项目(2009B050900007)；中央高校基本科研业务费专项资金(21609408)。

Brain derived neurotrophic factor promotes tube formation of cardiac microvascular endothelial cells

Liu Shao-kun1,2,3, Cao Liang1,2,3, Zhang Liang1,2,3, Shen Xiao-tao1,2,3, Zhao Bao-yin1,2,3, Qi Xu-feng1,2,3, Qin Jun-wen1,4, Cai Dong-qing1,2,3   

  1. 1Key Laboratory for Regenerative Medicine, Ministry of Education, 2Joint Laboratory for Regenerative Medicine, the Chinese University of Hong Kong-Jinan University, 3The International Base of Collaboration for Science & Technology (Jinan University), the Ministry of Science and Technology & Guangdong Province Science and Technology Committee, 4Institute of Reproductive Immunology, College of Life Science and Technology, Jinan University, Guangzhou  510632, Guangdong Province, China
  • Received:2010-12-23 Revised:2011-03-05 Online:2011-04-09 Published:2013-11-06
  • Contact: Cai Dong-qing, Professor, Doctoral supervisor, 1Key Laboratory for Regenerative Medicine, Ministry of Education, 2Joint Laboratory for Regenerative Medicine, the Chinese University of Hong Kong-Jinan University, 3The International Base of Collaboration for Science & Technology (Jinan University), the Ministry of Science and Technology & Guangdong Province, Jinan University, Guangzhou 510632, Guangdong Province, China tdongbme@jnu.edu.cn
  • About author:Liu Shao-kun★, Studying for master’s degree, 1Key Laboratory for Regenerative Medicine, Ministry of Education, 2Joint Laboratory for Regenerative Medicine, the Chinese University of Hong Kong-Jinan University, 3The International Base of Collaboration for Science & Technology (Jinan University), the Ministry of Science and Technology & Guangdong Province Science and Technology Committee, Jinan University, Guangzhou 510632, Guangdong Province Science and Technology Committee, China sakuelee@gmail.com
  • Supported by:

    the National 863 Program of China, No. 2007AA02Z105*; the National Natural Science Foundation of China, No. 30770886*, 30570369*, 30340038*, 30973158*; the Key Program of Guangdong Natural Science Foundation, No. 04105826*; Guangdong Grant for Science and Technology Development, No. 2004B30601007*; International Collaborated Grant of Guangdong, No. 2009B050900007*; the Fundamental Research Funds for the Central Universities, No. 21609408*

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摘要：

背景：研究发现脑源性神经营养因子可促进内皮细胞的存活，诱导血管新生，但其促进血管新生的细胞与分子机制尚不清楚。
目的：观察脑源性神经营养因子在3D体外管状形成模型中对心脏微血管内皮细胞管状结构形成的影响。
方法：分离提取大鼠心脏微血管内皮细胞并培养使之形成细胞小球，将心脏微血管内皮细胞小球接种于Ⅰ型胶原-甲基纤维素的3D环境中培养，然后分别加入50，70，100 μg/L的脑源性神经营养因子继续培养，于培养24，48 h观察并测量心脏微血管内皮细胞的分枝长度和分枝条数。
结果与结论：经分离的大鼠心脏微血管内皮细胞小球在Ⅰ型胶原-甲基纤维素3D环境中培养24 h，均可见管状分枝，且脑源性神经营养因子可促进管状分枝生长，以100 μg/L的脑源性神经营养因子的促生长效果最明显，分枝也最多。培养至    48 h，心脏微血管内皮细胞小球的分枝更长。说明脑源性神经营养因子在3D体外管状形成模型中可促进大鼠心脏微血管内皮细胞发芽及管状结构的形成，且具有剂量依赖性。

关键词: 心脏微血管内皮细胞, 脑源性神经营养因子, 血管新生, 管状结构形成

Abstract:

BACKGROUND: It has been discovered that brain derived neurotrophic factor (BDNF) can promote endothelial cell survival and induce angiogenesis. However, the cellular and molecular mechanism regarding to BDNF-induced angiogenesis is still unknown.
OBJECTIVE: To observe the effect of BDNF on tube formation of cardiac microvascular endothelial cells (CMECs) in three-dimensional (3D) in vitro tube formation assay.
METHODS: Rat CMECs were isolated and cultured until they formed a single spheroid. These spheroids were embedded in the mixture of collagen typeⅠ-carboxymethylcellulose (CMC). Then, 50 µg/L, 70 µg/L, 100 µg/L BDNF was added into the wells respectively. The number of branch and the cumulative length of all capillary-like structure were analyzed in 24 and 48 hours after BDNF treated respectively.
RESULTS AND CONCLUSION: The capillary-like structure for each CMECs spheroid was visible in 3D collagen typeⅠ-CMC containing after 24 hours culture. It was found that BDNF could promote tube formation of CMECs. The cumulative length and the branch of capillary-like structure of 100 µg/L BDNF group was significantly longer than that other groups respectively. The cumulative length of CMECs spheroids was significantly longer after 48 hours compared with at 24 hours. The results of this study suggested that BDNF could promote tube formation of CMECs in 3D in vitro tube formation assay. The promoting effect of BDNF is dosage dependence.

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