Key Laboratory of Medical Electrophysiology of Ministry of Education, Collaborative Innovation Center for Prevention and Treatment of Cardiovascular Disease of Sichuan Province, Drug Discovery Research Center, Southwest Medical University, Luzhou, China.
Laboratory for Cardiovascular Pharmacology, Department of Pharmacology, School of Pharmacy, Southwest Medical University, Luzhou, China.
Acta Physiol (Oxf). 2020 Sep;230(1):e13475. doi: 10.1111/apha.13475. Epub 2020 May 8.
Adipose-derived stem cells (ASCs) therapies are emerging as a promising approach to therapeutic angiogenesis. Therapeutic persistence and reduced primitive stem cell function following cell delivery remains a critical hurdle for the clinical translation of stem cells in current approaches.
Cultured ASCs were derived from subcutaneous white adipose tissue isolated from mice fed a normal diet (ND). Unilateral hindlimb ischaemia model was induced in high-fat diet (HFD)-fed mice by femoral artery interruption, after which photoactivated and non-light-treated ASCs were injected into the tail vein of mice. Laser Doppler imaging was conducted to measure the blood flow reperfusion. Capillary density was measured in the ischaemic gastrocnemius muscle. mRNA levels of angiogenic factors were determined by reverse-transcription polymerase chain reaction. Flow cytometry was used to determine the characterization of ASCs and endothelial progenitor cell (EPC). Human ASCs secretomes were analysed by liquid chromatography tandem mass spectrometry.
Our study demonstrated that photoactivated ND-ASCs prolonged functional blood flow perfusion and increased ASCs-derived EPC and neovascularization 38 days after ligation, when compared with saline-treated controls. Profiling analysis in ischaemic muscles showed upregulation of genes associated with pro-angiogenic factors after injection of photoactivated ND-ASCs when compared with the non-light-treated ASCs or saline treated HFD mice. Mass spectrometry revealed that light-treated ASCs conditioned medium retained a more complete pro-angiogenic activity with significant upregulation of angiogenesis related proteins.
Our data demonstrates that photoactivated ND-ASCs improve blood flow recovery and their injection may prove to be a useful strategy for the prevention and treatment of diabetic peripheral arterial disease.
脂肪来源干细胞（ASCs）疗法作为一种有前途的治疗血管生成方法正在出现。在目前的方法中，细胞递送后治疗的持久性和原始干细胞功能的降低仍然是干细胞临床转化的一个关键障碍。
从正常饮食（ND）喂养的小鼠的皮下白色脂肪组织中培养 ASC。通过股动脉阻断诱导高脂肪饮食（HFD）喂养的小鼠单侧后肢缺血模型，然后将光激活和非光照处理的 ASC 注射到小鼠尾静脉中。进行激光多普勒成像以测量血流再灌注。测量缺血比目鱼肌中的毛细血管密度。通过逆转录聚合酶链反应测定血管生成因子的 mRNA 水平。流式细胞术用于确定 ASC 和内皮祖细胞（EPC）的特征。通过液相色谱串联质谱分析人 ASC 分泌组。
我们的研究表明，与生理盐水处理的对照组相比，光激活的 ND-ASC 可延长功能血流灌注，并在结扎后 38 天增加 ASC 衍生的 EPC 和新血管形成。在缺血肌肉中的分析表明，与非光照 ASC 或生理盐水处理的 HFD 小鼠相比，注射光激活的 ND-ASC 后与促血管生成因子相关的基因上调。质谱分析显示，光照处理的 ASC 条件培养基保留了更完整的促血管生成活性，血管生成相关蛋白的显著上调。
我们的数据表明，光激活的 ND-ASC 可改善血流恢复，其注射可能被证明是预防和治疗糖尿病外周动脉疾病的有用策略。
