脑源性神经营养因子与心脏微血管内皮细胞的管状形成  

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

作　　者：刘少坤[1,2,3,4] 曹亮[1,2,3,4] 张靓[1,2,3,4] 沈晓涛[1,2,3,4] 赵宝寅[1,2,3,4] 齐绪峰[1,2,3,4] 秦俊文[1,5] 蔡冬青[1,2,3,4] 

机构地区：[1]暨南大学再生医学教育部重点实验室,广东省广州市510632 [2]暨南大学 [3]再生医学香港中文大学-暨南大学联合实验室,广东省广州市510632 [4]暨南大学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 [5]暨南大学生殖免疫研究所,广东省广州市510632

出　　处：《中国组织工程研究与临床康复》2011年第15期2769-2772,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：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)~~

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50,70,100μg/L的脑源性神经营养因子继续培养,于培养24,48h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24h,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100μg/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48h,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。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.

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 

分 类 号：R318[医药卫生—生物医学工程]