Comariţa Ioana Karla, Vîlcu Alexandra, Constantin Alina, Procopciuc Anastasia, Safciuc Florentina, Alexandru Nicoleta, Dragan Emanuel, Nemecz Miruna, Filippi Alexandru, Chiţoiu Leona, Gherghiceanu Mihaela, Georgescu Adriana
Institute of Cellular Biology and Pathology 'Nicolae Simionescu' of Romanian Academy, Bucharest, Romania.
'Carol Davila' University of Medicine and Pharmacy, Bucharest, Romania.
Front Cell Dev Biol. 2022 Mar 18;10:817180. doi: 10.3389/fcell.2022.817180. eCollection 2022.
Atherosclerosis is a progressive, chronic inflammatory disease of the large arteries caused by the constant accumulation of cholesterol, followed by endothelial dysfunction and vascular inflammation. We hypothesized that delivery of extracellular vesicles (EVs), recognized for their potential as therapeutic targets and tools, could restore vascular function in atherosclerosis. We explored by comparison the potential beneficial effects of EVs from subcutaneous adipose tissue stem cells (EVs (ADSCs)) or bone marrow mesenchymal stem cells (EVs (MSCs)) on the consequences of atherogenic diet on vascular health. Also, the influences of siRNA-targeting Smad2/3 (Smad2/3siRNA) on endothelial dysfunction and its key molecular players were analyzed. For this study, an animal model of atherosclerosis (HH) was transplanted with EVs (ADSCs) or EVs (MSCs) transfected or not with Smad2/3siRNA. For controls, healthy or HH animals were used. The results indicated that by comparison with the HH group, the treatment with EVs(ADSCs) or EVs(MSCs) alone or in combination with Smad2/3siRNA of HH animals induced a significant decrease in the main plasma parameters and a noticeable improvement in the structure and function of the thoracic aorta and carotid artery along with a decrease in the selected molecular and cellular targets mediating their changes in atherosclerosis: a decrease in expression of structural and inflammatory markers COL1A1, α-SMA, Cx43, VCAM-1, and MMP-2; a slight infiltration of total/M1 macrophages and T-cells; a reduced level of cytosolic ROS production; a significant diminution in plasma concentrations of TGF-β1 and Ang II proteins; significant structural and functional improvements (thinning of the arterial wall, increase of the inner diameter, enhanced distensibility, diminished VTI and Vel, and augmented contractile and relaxation responses); a reduced protein expression profile of Smad2/3, ATF-2, and NF-kBp50/p65 and a significant decrease in the expression levels of miR-21, miR-29a, miR-192, miR-200b, miR-210, and miR-146a. We can conclude that stem cell-derived EV therapies, especially the EVs (ADSCs) led to regression of structural and functional changes in the vascular wall and of key orchestrator expression in the atherosclerosis-induced endothelial dysfunction; transfection of EVs with Smad2/3siRNA amplified the ability of EVs(ADSCs) or EVs(MSCs) to regress the inflammation-mediated atherosclerotic process.
动脉粥样硬化是一种由胆固醇持续积累引发的大动脉进行性慢性炎症性疾病，随后会出现内皮功能障碍和血管炎症。我们推测，因其作为治疗靶点和工具的潜力而受到认可的细胞外囊泡（EVs），可能会恢复动脉粥样硬化中的血管功能。我们通过比较来自皮下脂肪组织干细胞的细胞外囊泡（EVs(ADSCs)）或骨髓间充质干细胞的细胞外囊泡（EVs(MSCs)）对致动脉粥样硬化饮食对血管健康影响的潜在有益作用。此外，还分析了靶向Smad2/3的小干扰RNA（Smad2/3siRNA）对内皮功能障碍及其关键分子参与者的影响。在本研究中，将动脉粥样硬化动物模型（HH）移植了用或未用Smad2/3siRNA转染的EVs(ADSCs)或EVs(MSCs)。作为对照，使用了健康或HH动物。结果表明，与HH组相比，单独使用或与Smad2/3siRNA联合使用EVs(ADSCs)或EVs(MSCs)治疗HH动物，可使主要血浆参数显著降低，胸主动脉和颈动脉的结构和功能有明显改善，同时介导动脉粥样硬化变化的选定分子和细胞靶点减少：结构和炎症标志物COL1A1、α-SMA、Cx43、VCAM-1和MMP-2的表达降低；总/M1巨噬细胞和T细胞的轻微浸润；胞质活性氧生成水平降低；血浆中TGF-β1和Ang II蛋白浓度显著降低；结构和功能显著改善（动脉壁变薄、内径增加、扩张性增强、VTI和Vel降低以及收缩和舒张反应增强）；Smad2/3、ATF-2和NF-kBp50/p65的蛋白表达谱降低，miR-21、miR-29a、miR-192、miR-200b、miR-210和miR-146a的表达水平显著降低。我们可以得出结论，干细胞衍生的EV疗法，尤其是EVs(ADSCs)，可导致血管壁结构和功能变化以及动脉粥样硬化诱导的内皮功能障碍中关键协调因子表达的消退；用Smad2/3siRNA转染EVs可增强EVs(ADSCs)或EVs(MSCs)消退炎症介导的动脉粥样硬化过程的能力。
