中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (16): 4077-4087.doi: 10.12307/2026.679

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

音猬因子信号通路调控小鼠胚胎心脏流出道的形成与分隔

姚凯宁，闫玉楠，周祎凡，师  亮，曹锡梅，Zeeshan Rahim，杨艳萍   

  1. 山西医科大学组织胚胎学教研室，山西省太原市  030001
  • 收稿日期:2025-04-27 接受日期:2025-08-19 出版日期:2026-06-08 发布日期:2025-11-26
  • 通讯作者: 杨艳萍，博士，教授，山西医科大学组织学与胚胎学教研室，山西省太原市 030001
  • 作者简介:姚凯宁，女，2000年生，河北省唐山市人，汉族，山西医科大学在读硕士，主要从事胚胎心脏发育研究。
  • 基金资助:
    山西省科技厅应用基础研究计划面上项目(202303021221135)，项目负责人：杨艳萍；山西省科技厅应用基础研究计划面上项目(202303021211247)，项目负责人：师亮；山西省研究生教育创新计划(2023JG085)，项目负责人：师亮；山西省高等学校教学改革创新项目(J20230486)，项目负责人：师亮

Sonic hedgehog signaling pathway regulates the formation and septation of the outflow tract in the embryoic mouse heart

Yao Kaining, Yan Yunan, Zhou Yifan, Shi Liang, Cao Ximei, Zeeshan Rahim, Yang Yanping   

  1. Department of Histology and Embryology, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • Received:2025-04-27 Accepted:2025-08-19 Online:2026-06-08 Published:2025-11-26
  • Contact: Yang Yanping, PhD, Professor, Department of Histology and Embryology, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • About author:Yao Kaining, MS candidate, Department of Histology and Embryology, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • Supported by:
    Applied Basic Research Program of Department of Science and Technology of Shanxi Province (General Programs), No. 202303021221135 (to YYP); Applied Basic Research Program of Department of Science and Technology of Shanxi Province (General Programs), No. 202303021211247 (to SL); Graduate Education Innovation Program of Shanxi Province, No. 2023JG085 (to SL); Shanxi Province Higher School Teaching Reform Innovation Program, No. J20230486 (to SL)

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摘要：




文题释义：
第二生心区：由咽前间充质构成。第二生心区的祖细胞增殖、迁移，分化为右心室、流出道、心房心肌细胞及大动脉平滑肌细胞。
音猬因子信号通路：由音猬因子配体、跨膜蛋白受体Ptch家族(Ptch1和Ptch2)、跨膜蛋白Smoothened、核转录因子Gli蛋白、氟尿嘧啶抑制剂及下游靶基因构成。音猬因子信号通路参与脊椎动物的多种生物学过程，包括胚胎心肺发育、细胞生长分化等。

背景：第二生心区及心脏神经嵴共同参与流出道的形成与分隔，但音猬因子信号通路在该过程中调控作用的确切机制尚未明确。
目的：探究第二生心区形成过程中音猬因子信号通路各组分的调控作用，确认音猬因子信号通路在流出道形成与分隔过程中的调控作用。
方法：从妊娠ICR小鼠中获取胚龄10-15 d小鼠胚胎作为实验材料，经石蜡包埋后制备连续切片样本，通过免疫组化染色、免疫荧光双染实验观察胰岛素基因增强结合蛋白1、音猬因子、Ptch1、Ptch2、Smoothened、Gli3及骨形态发生蛋白2的时空表达模式；剥离10.5 d胚龄小鼠胚胎的前肠与相邻间充质，通过蛋白印迹分析与免疫共沉淀实验探究音猬因子信号通路与骨形态发生蛋白和第二生心区细胞与心脏神经嵴细胞之间的关系。 
结果与结论：①在胚龄10-11.5 d小鼠胚胎中，音猬因子与Gli3的表达见于咽内胚层，而Ptch1、Ptch2与Smoothened表达与呼吸内胚层的发生相偶联；呼吸内胚层形成导致胰岛素基因增强结合蛋白1阳性第二生心区祖细胞增多并参与流出道的发育。音猬因子信号通路各组分未在鳃弓核心间充质表达，但出现于心包腔背侧壁。②在流出道心内膜垫形成与融合过程中，Ptch1、Ptch2与Smoothened表达于心肌，Gli3表达于心肌与心内膜垫的间充质细胞、主动脉与肺动脉干管壁。③在胚龄胚龄10.5-11 d小鼠胚胎中，胰岛素基因增强结合蛋白1和骨形态发生蛋白2的共表达分布于前肠内胚层及第二生心区；沿弓动脉分布的激活增强子结合蛋白2α阳性心脏神经嵴细胞同时表达Gli3或骨形态发生蛋白2，免疫共沉淀显示激活增强子结合蛋白2α与Gli3或骨形态发生蛋白2存在相互作用。④结果表明，音猬因子信号通路参与第二生心区发育，并且在第二生心区不同亚群的形成、迁移与分化过程中发挥不同作用；音猬因子信号通路在流出道心内膜垫形成与融合过程中持续产生作用；Gli3、骨形态发生蛋白2共同调节心脏神经嵴细胞的迁移。

https://orcid.org/0009-0007-1068-9381(姚凯宁)


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词:

心脏流出道, 第二生心区, 心脏神经嵴, 音猬因子信号通路, 骨形态发生蛋白2, 激活增强子结合蛋白2α

Abstract: BACKGROUND: Second heart field and cardiac neural crest are involved in the formation and septation of the outflow tract. However, the exact regulatory mechanism of the Sonic hedgehog (Shh) signaling pathway in this process have not been clarified.
OBJECTIVE: To explore the regulatory role of the components of the Shh signaling pathway in the formation and septation of outflow tract.
METHODS: Mouse embryos at embryonic 10-15 days were obtained from pregnant ICR mice, and serial section samples were prepared after paraffin embedding to observe the spatiotemporal expression patterns of insulin gene enhancer binding protein-1 (isl-1), sonic hedgehog (Shh), Ptch1, Ptch2, Smoothened, Gli3, and bone morphogenetic protein 2 by immunohistochemical staining and double immunofluorescence staining. The foregut and adjacent mesoderm mesenchyme of mouse embryos at embryonic 10.5 days were dissected and the relationship between the Shh signaling pathway and bone morphogenetic proteins and cells in the second heart field and cardiac neural crest was explored by western blot and co-immunoprecipitation.
RESULTS AND CONCLUSION: (1) On embryonic 10-11.5 days, the expression of Shh and Gli3 was seen in the pharyngeal endoderm, whereas the expression of Ptch1, Ptch2, and Smoothened was coupled to the development of respiratory endoderm. The respiratory endoderm formation resulted in an increase in isl-1-
positive progenitors in the second heart field and was involved in the development of the outflow tract. Components of the Shh signaling pathway were not expressed in the core mesenchyme of the branchial arch, but appeared in the dorsal wall of the pericardial cavity. (2) During the formation and fusion of the outflow tract endocardial cushion, Ptch1, Ptch2, and Smoothened were expressed in the myocardium, and Gli3 was expressed in the mesenchymal cells of the myocardium and endocardial cushion, as well as in the walls of the aortic and pulmonary trunk. (3) On embryonic 10-11.5 days, isl-1 and bone morphogenetic protein 2 were co-expressed in the foregut endoderm and second heart field. Activating protein 2α-positive cardiac neural crest cells surrounding the aortic arch arteries co-expressed either Gli3 or bone morphogenetic protein 2. Co-immunoprecipitation result showed that activating protein 2α interacted with Gli3 or bone morphogenetic protein 2. These findings indicate that the Shh signaling pathway contributes to second heart field development, and plays variant roles in the formation, migration, and differentiation of the second heart field subpopulations. The Shh signaling pathway has a sustained role in endocardial cushion formation and fusion of the outflow tract. Gli3 and bone morphogenetic protein 2 cooperatively regulate the migration of cardiac neural crest cells.


Key words: cardiac outflow tract, second heart field, cardiac neural crest, sonic hedgehog pathway, bone morphogenetic protein 2, activating enhancer binding protein 2α

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