Laboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil; National Institute of Science and Technology for Regenerative Medicine, Rio de Janeiro, Brazil.
Laboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
Life Sci. 2023 Sep 15;329:121988. doi: 10.1016/j.lfs.2023.121988. Epub 2023 Jul 29.
To evaluate BM-MSCs and their extracellular vesicles (EVs) preconditioned with hypoxia or normoxia in experimental pulmonary arterial hypertension (PAH).
BM-MSCs were isolated and cultured under normoxia (MSC-N, 21%O) or hypoxia (MSC-H, 1%O) for 48 h. EVs were then isolated from MSCs under normoxia (EV-N) or hypoxia (EV-H). PAH was induced in male Wistar rats (n = 35) with monocrotaline (60 mg/kg); control animals (CTRL, n = 7) were treated with saline. On day 14, PAH animals received MSCs or EVs under normoxia or hypoxia, intravenously (n = 7/group). On day 28, right ventricular systolic pressure (RVSP), pulmonary acceleration time (PAT)/pulmonary ejection time (PET), and right ventricular hypertrophy (RVH) index were evaluated. Perivascular collagen content, vascular wall thickness, and endothelium-mesenchymal transition were analyzed.
PAT/PET was lower in the PAH group (0.26 ± 0.02, P < 0.001) than in CTRLs (0.43 ± 0.02) and only increased in the EV-H group (0.33 ± 0.03, P = 0.014). MSC-N (32 ± 6 mmHg, P = 0.036), MSC-H (31 ± 3 mmHg, P = 0.019), EV-N (27 ± 4 mmHg, P < 0.001), and EV-H (26 ± 5 mmHg, P < 0.001) reduced RVSP compared with the PAH group (39 ± 4 mmHg). RVH was higher in the PAH group than in CTRL and reduced after all therapies. All therapies decreased perivascular collagen fiber content, vascular wall thickness, and the expression of endothelial markers remained unaltered; only MSC-H and EV-H decreased expression of mesenchymal markers in pulmonary arterioles.
MSCs and EVs, under normoxia or hypoxia, reduced right ventricular hypertrophy, perivascular collagen, and vessel wall thickness. Under hypoxia, MSCs and EVs were more effective at improving endothelial to mesenchymal transition in experimental PAH.
评估缺氧或常氧预处理的骨髓间充质干细胞（BM-MSCs）及其细胞外囊泡（EVs）在实验性肺动脉高压（PAH）中的作用。
将 BM-MSCs 在常氧（MSC-N，21%O）或缺氧（MSC-H，1%O）条件下培养 48 小时。然后从常氧（EV-N）或缺氧（EV-H）下的 MSC 中分离 EVs。雄性 Wistar 大鼠（n=35）用单克隆旋毛虫（60mg/kg）诱导 PAH；对照组（CTRL，n=7）用生理盐水处理。第 14 天，PAH 动物静脉内接受常氧或缺氧条件下的 MSC 或 EV（n=7/组）。第 28 天，评估右心室收缩压（RVSP）、肺动脉加速时间（PAT）/肺动脉射血时间（PET）和右心室肥厚（RVH）指数。分析血管周围胶原含量、血管壁厚度和内皮-间充质转化。
PAH 组的 PAT/PET 低于对照组（0.26±0.02，P<0.001），仅 EV-H 组增加（0.33±0.03，P=0.014）。MSC-N（32±6mmHg，P=0.036）、MSC-H（31±3mmHg，P=0.019）、EV-N（27±4mmHg，P<0.001）和 EV-H（26±5mmHg，P<0.001）均降低 RVSP ，与 PAH 组（39±4mmHg）相比。PAH 组的 RVH 高于对照组，并在所有治疗后降低。所有治疗均降低血管周围胶原纤维含量、血管壁厚度，内皮标志物的表达保持不变；只有 MSC-H 和 EV-H 降低了肺小动脉中间充质标志物的表达。
常氧或缺氧条件下的 MSC 和 EV 可减轻右心室肥厚、血管周围胶原和血管壁厚度。在缺氧条件下，MSC 和 EV 在改善实验性 PAH 中的内皮-间充质转化方面更有效。
