学术
Sonichedgehog信号通路对AGM区来源的成血-血管细胞增殖、迁移和分化作用
付劲蓉 刘文励 周剑锋 剥、汉英 冉丹 郑邈 罗莉 张恒 周玉峰
武汉,华中科技大学同济医学院附属同济医院血液科,430030
摘 要:
目的探讨Sonichedgehog（Shh）信号通路对小鼠主动脉-中肾-肾上腺（AGM）区来源的成血．血管细胞增殖、分化和迁移的影响。方法联合应用抗小鼠CD34、Flkl单克隆抗体（单抗）的磁珠，从孕11dBALB／c鼠胚胎AGM区分离培养成血-血管干细胞，并同时培养AGM区来源基质细胞，用流式细胞术对所分离细胞进行表型鉴定。用免疫组织化学法检测基质细胞表达Shh蛋白的情况；借助TransweU细胞共培养体系，观察在AGM的基质细胞共培养体系中，不同浓度的Shh活性肽及其单抗对成血．血管干细胞增殖、细胞周期、迁移及向内皮细胞定向分化的影响。结果AGM区基质细胞高表达Shh蛋白（阳性率高达90％以上），并可促进成血-血管干细胞增殖，其增殖效应可被10．00μg／mlShh单抗阻断；0．10-10．00μg／ml外源性Shh活性蛋白与AGM区基质细胞共培养的成血-血管干细胞，可使成血．血管干细胞长时间增殖，并促进细胞迁移，但并不诱导细胞凋亡与分化；而当0．10-10．00μ／ml外源性Shh活性蛋白作用于单独培养的成血-血管干细胞，细胞经过短时间增殖后，随后发生凋亡并向内皮细胞分化。结论Shh信号通路参与了成血-血管干细胞的增殖、分化、凋亡及迁移等生物学特性的调控，但其具体作用存在时空的差异，AGM区微环境对Shh信号通路具有整合和调试的作用。[著者文摘]
文章出处:
《中华血液学杂志》-2007年28卷11期 -757-761页
栏目信息:
分 类 号:
Role of the sonic hedgehog pathway in regulation of the proliferation, migration and differentiation of hemangiobiast derived from AGM
FU Jin-rong, LIU Wen-li, ZHOU Jian-feng, SUN Han-ying, RAN Dan, ZHENG Miao, LUO Li, ZHANG Heng, ZHOU Yu-feng. （Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China）
Abstract:
Objective To explore the role of sonic hedgehog （Shh） pathway in regulating the prolif- eration, migration and differentiation of hemangioblasts derived from aorta-gonad-mesonephros （AGM）. Methods The hemangioblasts were isolated from AGM region of 11-day postcoitum （dpc） murine embryos by using the immuno-magnetic beads cooted with CD34 and Flkl monoclonal antibodies. The phenotypic ana- lysis of henmngioblasts and AGM-derived stromal ceils were detected by flow cytometry. The secretion of Shh was examined by immunohistochemical staining. The roles of Shh in regulating the proliferation, migration and differentiation of hemangioblasts in the transweil non-contact coculture system with AGM-derived stromal ceils were observed by adding exogenous Shh N-Terminus and its antibody. Results The protein of Shh was highly expressed on AGM-derived stromal ceils. The proliferation of hemangioblasts was promoted when co-cultured with AGM-derived stromal ceils, and the effects of the latter could be blocked by antibody of Shh. The prolif- eration of hemangioblasts was strengthened further and kept for a long time without differentation and apoptosis when exogenous Shh N-Terminus was added into the transweil non-contact co-culture system with AGM-de- rived stromal ceils. When exogenous Shh N-Terminus was added into the cultural supernatant of hemangio- blasts without AGM-derived stromal ceils, the hemangioblasts were observed to be induced to apoptosis or dif- ferentiation after a short time of proliferation. Furethmore, the ability of migration could be promoted in the co-cultured hemangioblasts by adding exogenous Shh N-Terminus. Conclusion Shh pathway probably in- volves in the regulation of the proliferation, differentiation, apoptosis and migration of hemangioblasts, and is regulated by the AGM microenvironment.[著者文摘]
Key words:
Sonic hedgehog; Hemangioblast; Proliferation; Differentiation ; Migration
基金资助:
国家自然科学基金资助项目（30570773）
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