骨形态发生蛋白4/DNA结合抑制剂2信号通路在晚期内皮祖细胞修复血管内皮损伤中的作用  

Bone morphogenetic protein 4/inhibitor of DNA binding 2 signaling pathway regulates late outgrowth endothelial progenitor cell-mediated endothelial injury repair

作　　者：梁建文 苏晨[2] 陈龙[2] 夏文豪[2] 陶军[2] LIANG Jian-wen;SU Chen;CHEN Long;XIA Wen-hao;TAO Jun Cardiology(Department,The Eighth Affiliated Hospital of Sun Yat-sen University,Guangdong Shenzhen 518000,China)

机构地区：[1]中山大学附属第八医院心血管内科,广东深圳518000 [2]中山大学附属第一医院心血管内科

出　　处：《中华高血压杂志》2018年第8期759-765,共7页Chinese Journal of Hypertension

摘　　要：目的研究骨形态发生蛋白4(BMP4)信号通路在晚期内皮祖细胞(LEPC)介导的血管内皮修复中的作用与分子机制。方法分离健康人外周血单核细胞体外培养7d和28d分别得到早期内皮祖细胞(EEPC)和LEPC,观察BMP4在EEPC和LEPC中的表达情况。体外采用短发卡RNA(shRNA)沉默技术和基因转染干预BMP4的表达,观察其下游分子靶点DNA结合抑制剂2(Id2)表达情况及对LEPC的迁移、黏附等功能的影响。建立裸鼠颈动脉内膜拉脱损伤模型,在体观察干预BMP4信号通路如何介导LEPC修复血管内皮损伤。结果相对于EEPC,BMP4选择性高表达于LEPC(P<0.01)。shRNA法沉默LEPC的BMP4表达后,其下游靶点Id2的表达水平下调(P<0.01);相应的LEPC体外迁移、黏附能力明显减低(P<0.01);将沉默了BMP4的LEPC在体移植到裸鼠后其颈动脉内皮损伤的修复面积明显减小[(34±6)%比(58±7)%,P<0.01]。基因转染上调LEPC的BMP4表达后,其下游的Id2表达明显增强(P<0.01),LEPC的迁移、黏附功能改善(P<0.01),移植到裸鼠体内后其颈动脉内皮损伤的修复面积明显增加[(75±5)%比(59±6)%,P<0.01]。结论LEPC修复血管内皮损伤的作用与BMP4/Id2信号通路的表达密切相关。Objective To investigate the effect of bone morphogenetic protein 4（BMP4）-modified signaling pathway on human late outgrowth endothelial progenitor cells（LEPC）-related endothelial repair and the underlying molecular mechanism. Methods In vitro,after a 7-day and 28-day culture,BMP4 expression was observed in human early endothelial progenitor cells（EEPC）and LEPC. After inhibiting BMP4 with short hairpin RNA（shRNA）-mediated knockdown or up-regulating BMP4 with gene transfer,the Id2 expression and LEPC capacities of migration and adhesion were investigated in vitro. In vivo endothelial repair ability of BMP4 induced LEPC-related endothelial repair in nude mice with carotid artery denudation injury was also examined. Results BMP4 expressed selectively much higher in LEPC than in EEPC（P〈0.01）. Moreover,inhibition of BMP4 with shRNA down-regulated BMP4 and Id2 expression（P〈0.01）,impaired in vitro LEPC capacities（P〈0.01）and decreased the repaired areas of injured carotid artery endothelial in nude mice[（34±6）% vs（58±7）%,P〈0.01]. Furthermore,BMP4 gene transfer remarkably activated BMP4-mediated signaling pathway,resulting in increasing Id2 expression（P〈0.01）,facilitating therapeutic endothelial repair capacity of LEPC in nude mice with carotid artery denudation injury[（75±5）%vs（59±6）%,P〈0.01]. Conclusion BMP4-related signaling pathway is essential for endothelial repair capacity of LEPC in human.

关 键 词：内皮损伤 晚期内皮祖细胞 骨形态发生蛋白4/DNA结合抑制剂2信号通路 内皮修复 细胞治疗 

分 类 号：R54[医药卫生—心血管疾病]