The JAK/STAT3 signalling pathway regulated angiogenesis in an endothelial cell/adipose-derived stromal cell co- culture,3D gel model
The JAK/STAT3 signalling pathway regulated angiogenesis in an endothelial cell/adipose-derived stromal cell co- culture,3D gel model
复制标题
JAK/STAT3 信号通路调节内皮细胞/脂肪源性基质细胞共培养 3D 凝胶模型中的血管生成。
DOI：
10.1111/cpr.12307
复制
发表时间：
2017-02-01
影响因子：
8.5
通讯作者：
Cai, Xiaoxiao
中科院分区：
文献类型：
--
作者：
Xue, Changyue;Xie, Jiamin;Cai, Xiaoxiao
目的：本研究的目的是探讨JAK/STAT 3信号通路在血管生成中的作用。材料和方法：体外建立的模型，涉及三维胶原凝胶植入内皮细胞（ECs）从红色荧光蛋白标记的小鼠，和脂肪来源的基质细胞（ASCs）从绿色荧光蛋白标记的小鼠。用激光共聚焦显微镜观察JAK/STAT 3通路抑制剂和激活剂处理后血管生成现象。结果：JAK/STAT 3通路抑制剂Stattic可抑制血管生成，但其抑制血管生成的作用机制尚不清楚。我们发现VEGFA和cyclin D1的蛋白水平受JAK/STAT 3通路的调节，流式细胞术进一步证实了EC和ASCs细胞周期参数的变化。Stattic可下调内皮细胞中VEGFA/B、VEGFR 2、MMP-2、MMP-9、IGF-1和b-FGF基因的表达，而奥氮平可显著上调这些基因的mRNA水平。结论：JAK/STAT 3信号通路参与了胶原凝胶中血管生成的调控，其调控机制包括血管长度、直径和芽数的变化。JAK/STAT 3信号通路通过调节血管生成相关生长因子的数量和直接调节细胞周期来调节血管生成。
Objectives: The aim of the study was to investigate the role of the JAK/STAT3 signalling pathway in angiogenesis.Materials and methods: The model established in vitro, involved a 3D collagen gel being implanted with endothelial cells (ECs) from red fluorescent protein-labelled mice, and adipose-derived stromal cells (ASCs) from green fluorescent protein-labelled mice. Phenomena of angiogenesis, after treatment by the inhibitor and the activator of JAK/STAT3 pathway respectively, were observed using confocal laser scanning microscopy. Transwell co-culture of ECs and ASCs was used to elucidate mechanisms.Results: Stattic, inhibitor of JAK/STAT3 pathway, attenuated angiogenesis in the model. In contrast, angiogenesis was promoted after treatment of Olanzapine, an activator.We found that protein levels of VEGFA and cyclin D1 were regulated by the JAK/STAT3 pathway, and flow cytometry further confirmed variations in cell cycle parameters of ECs and ASCs. Genes VEGFA/B, VEGFR2, MMP-2, MMP-9, IGF-1 and b-FGF were down-regulated by Stattic in ECs, while Olanzapine significantly up-regulated mRNA levels of these genes. As for ASCs, genes VEGFA, MMP-2, MMP-9, IGF-1 and b-FGF were modulated by the JAK/STAT3 pathway.Conclusions: Angiogenesis in the 3D collagen gel was regulated by the JAK/STAT3 pathway which involved changes in vessel length, vessel diameter and sprout number. The underlying mechanism was that the JAK/STAT3 signalling pathway regulated angiogenesis by modulation of numbers of angiogenesis-related growth factors and by direct regulation of cell cycle.