通过慢病毒介导的短发夹RNA对成纤维细胞的转化生长因子β(TGF-β)信号通路进行干扰,采用双光子荧光显微成像技术并结合基于细胞-细胞外基质三维系统的图像信息学方法,研究了植于多孔胶原蛋白支架中成纤维细胞分化过程的TGF-β信号通路.结果表明,在介导成纤维细胞分化的过程中,生长因子TGF-β1比TGF-β3发挥的作用更大,在TGF-β1介导的成纤维细胞分化过程中,蛋白SMAD1和SMAD3通过不同方式也发挥了重要作用.所得结果与通过生化或遗传学方法的结果相吻合.
Stable protein expression interference of key components of transforming growth factor(TGF)-βpathway in fibroblasts was achieved by transfection with a shRNA lentivirus.Two-photon microscopy,combined with an image informatics framework that suitable for 3Dcell-matrix system,was used to study the TGF-βsignaling in fibroblasts inside porous collagen scaffolds.It is concluded that,compared with TGF-β3,TGF-β1plays a more important role in mediating myofibroblast differentiation,and that SMAD1 and SMAD3play key roles in TGF-β1mediated myofibroblast differentiation but in different ways.These conclusions were consistent with the results obtained by using biochemical or genetic methods.