ADSC通过SDF1-RhoA/ROCK通路调控巨噬细胞募集促进缺血肌肉血管新生的作用机制研究

脂肪干细胞（ADSC）移植可促进严重缺血肢体（CLI）血管新生、改善患肢预后，但其作用机制尚未阐明。研究表明巨噬细胞（Mφ）浸润和表型转化在缺血肌肉血管新生中起重要调控作用。我们前期研究发现，ADSC移植可显著增加缺血肌肉中Mφ数量，且在体外ADSC可显著增强Mφ的迁移作用，而阻断SDF1或沉默PI3K/RhoA均可抑制该过程。据此我们假设，ADSC通过SDF1/PI3K介导的RhoA/ROCK通路调控Mφ向缺血肌肉内募集，进而通过表型转化促进缺血肢体血管新生。在前期工作基础上，拟在体内、外进一步研究ADSC对Mφ迁移的影响，通过基因转染、RNA干扰等探索SDF1-RhoA/ROCK通路在ADSC调控Mφ迁移中的作用，并结合ApoE基因敲除小鼠CLI模型，运用巨噬细胞清除、体内示踪等方法明确ADSC调控Mφ募集在缺血肌肉血管新生中的作用和机制，为CLI的干细胞治疗提供新的思路和理论依据。

The transplantation of adipose-derived stem cell (ADSC) can promote angiogenesis of critical ischemic limb (CLI) and improve the prognosis of affected limb. Studies have found that macrophage (Mφ) infiltration and phenotypic transformation play an important role in ischemic muscle angiogenesis, but the specific regulatory mechanism is not clear. Our previous studies found that ADSC transplantation significantly increased the number of Mφ in ischemic muscle. In vitro experiments also confirmed that ADSC could significantly enhance the migration of Mφ. However, suppressing SDF1 or silencing PI3K/RhoA could inhibit this effect. Therefore, we hypothesized that ADSC could regulate the recruitment of Mφ into ischemic muscles through SDF1/PI3K-mediated RhoA/ROCK pathway, and then promote angiogenesis in ischemic limbs through phenotypic transformation. This study intends to further study the influence of ADSC on Mφ migration in vivo and vitro on the basis of previous work and explore the role of SDF1-RhoA/ROCK pathway on Mφ migration mediated by ADSC through gene transfection, RNA interference, etc. In combination with ApoE knockout mouse CLI model, macrophage-clearing technique and in vivo-tracing method would be used to study the role and mechanism of ADSC in inducing Mφ recruitment and promoting angiogenesis in ischemic muscles, so as to provide new ideas and theoretical basis for stem cell therapy of CLI.