N-acetylcysteine differentially regulates the populations of bone marrow and circulating endothelial progenitor cells in mice with limb ischemia
N-acetylcysteine differentially regulates the populations of bone marrow and circulating endothelial progenitor cells in mice with limb ischemia
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DOI：
10.1016/j.ejphar.2020.173233
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发表时间：
2020-08-15
影响因子：
5
通讯作者：
Liu, Zhenguo
中科院分区：
文献类型：
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作者：
Cui, Yuqi;Liu, Lingjuan;Liu, Zhenguo
内皮祖细胞（Endothelial progenitor cells，EPCs）是组织修复和再生的重要细胞，尤其是在缺血性损伤后，其表型和生物学特性具有很大的异质性。活性氧参与调节EPC的数量和功能。N-乙酰半胱氨酸（NAC）抑制缺血诱导的活性氧形成，促进缺血肢体恢复。本研究旨在探讨NAC对肢体缺血小鼠骨髓和血液中EPC亚群的影响。通过结扎雄性C57 BL/6小鼠的股动脉诱导肢体缺血，有或没有NAC治疗。在基线、结扎后第3天（急性缺血）和第21天（慢性）分析BM和血细胞中的EPC亚群、细胞内活性氧产生、细胞增殖和凋亡。使用c-Kit+/CD 31+、Sca-1+/Flk-1+、CD 34 +/CD 133+和CD 34 +/Flk-1+来定义EPC亚群。分别于缺血后第3、7、14和21天采用激光多普勒灌注成像、平板运动试验和免疫组织化学方法评价肢体血流、功能、肌肉结构和毛细血管密度。在急性和慢性缺血小鼠的BM和血液中，循环和BM单核细胞和EPCs群体中的活性氧产生显著增加。NAC治疗有效地阻断了缺血诱导的循环和BM单核细胞中活性氧的产生，并选择性地增加了循环中的EPC群体，而不是BM，在慢性缺血小鼠中保持了增殖，并增强了肢体血流和功能恢复，同时防止了急性缺血诱导的BM和循环EPC的增加。这些数据表明，NAC选择性地增强慢性肢体缺血小鼠循环EPC群体。
Endothelial progenitor cells (EPCs) are important to tissue repair and regeneration especially after ischemic injury, and very heterogeneous in phenotypes and biological features. Reactive oxygen species are involved in regulating EPC number and function. N-acetylcysteine (NAC) inhibits ischemia-induced reactive oxygen species formation and promotes ischemic limb recovery. This study was to evaluate the effect of NAC on EPC subpopulations in bone marrow (BM) and blood in mice with limb ischemia. Limb ischemia was induced by femoral artery ligation in male C57BL/6 mice with or without NAC treatment. EPC subpopulations, intracellular reactive oxygen species production, cell proliferation and apoptosis in BM and blood cells were analyzed at baseline, day 3 (acute ischemia) and 21 (chronic) after ligation. c-Kit+/CD31+,Sca-1+/Flk-1+,CD34+/CD133+, and CD34+/Flk-1+ were used to define EPC subpopulations. Limb blood flow, function, muscle structure, and capillary density were evaluated with laser Doppler perfusion imaging, treadmill test, and immunohistochemistry, respectively, at day 3, 7, 14 and 21 post ischemia. Reactive oxygen species production in circulating and BM mononuclear cells and EPCs populations were significantly increased in BM and blood in mice with acute and chronic ischemia. NAC treatment effectively blocked ischemia-induced reactive oxygen species production in circulating and BM mononuclear cells, and selectively increased EPC population in circulation, not BM, with preserved proliferation in mice with chronic ischemia, and enhanced limb blood flow and function recovery, while preventing acute ischemia-induced increase in BM and circulating EPCs. These data demonstrated that NAC selectively enhanced circulating EPC population in mice with chronic limb ischemia.