Hypoxia-Induced Mesenchymal Stromal Cells Exhibit an Enhanced Therapeutic Effect on Radiation-Induced Lung Injury in Mice due to an Increased Proliferation Potential and Enhanced Antioxidant Ability
Hypoxia-Induced Mesenchymal Stromal Cells Exhibit an Enhanced Therapeutic Effect on Radiation-Induced Lung Injury in Mice due to an Increased Proliferation Potential and Enhanced Antioxidant Ability
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缺氧诱导的间充质干细胞由于增殖潜力增加和抗氧化能力增强，对小鼠辐射引起的肺损伤具有增强的治疗作用
DOI：
10.1159/000485490
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Yang, Yanyong
中科院分区：
文献类型：
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作者：
Li, Bailong;Li, Cheng;Yang, Yanyong
背景/目标：放射治疗是胸部肿瘤的重要治疗方法，然而，伴随放射治疗的副作用导致肿瘤控制有限和患者生活质量下降。在这些副作用中，放射性肺损伤（RILI）是最严重和最常见的。因此，需要对RILI进行有效的补救。间充质干细胞（Mesenchymal stromal cells，MSCs）是一种多能成体干细胞，已被证明是一种有效的治疗由组织损伤引起的疾病的方法。然而，与其他损伤不同，RILI由于局部缺氧和照射肺中广泛的活性氧（ROS）而从植入的MSC获得有限的治疗效果。由于MSC的存活率差主要是由于缺氧和ROS的产生，我们假设持续和适应性缺氧处理诱导植入MSC对缺氧应激的抵抗力增强。本研究的目的是调查是否持续和适应性低氧治疗的骨髓间充质干细胞移植前在受伤的小鼠提高生存和改善RILI的疗效。研究方法：从6周龄雄性C57 BL 6/J小鼠的骨髓中获得原代bmMSCs，并在常氧条件（21%O-2）或缺氧条件（2.5%O-2）下培养。小鼠胸部照射（20戈伊）后注射常氧/缺氧MSC。采用HE染色、Masson染色、α-SMA染色等方法观察MSCs对RILI的治疗作用，同时采用B. Li，C. Li，M. Zhu和Y。张对这项工作同样作出了贡献。使用ELISA测量炎性因子。使用显微镜记录体外MSC的形态并通过流式细胞术鉴定，使用CCK-8测定法测量细胞活力，通过EdU测定法检测增殖潜力，并使用ROS荧光探针测量ROS水平。此外，还通过Western blotting检测了HIF-1 α和几种存活途径蛋白（Akt、p-Akt、Caspase-3）。结果：骨髓间充质干细胞移植可减轻早期放射性肺炎和晚期肺纤维化。然而，缺氧MSC显示出更明显的治疗效果相比，常氧MSC。与常氧MSC相比，缺氧MSC表现出更大的细胞活力，增强的增殖潜力，降低的ROS水平和增加的耐缺氧和ROS应激性。此外，低氧MSC实现了更高水平的HIF-1 α和Akt的活化，并且HIF-1 α在耐药性的发展中起关键作用。结论：低氧通过促进间充质干细胞增殖和提高其抗氧化能力增强间充质干细胞对放射性肺损伤的治疗作用，HIF-1 α介导。(c)2017作者（s）由S. Karger AG，巴塞尔
Background/Aims: Radiation therapy is an important treatment for thoracic cancer; however, side effects accompanied with radiotherapy lead to limited tumor control and a decline in patient quality of life. Among these side effects, radiation-induced lung injury (RILI) is the most serious and common. Hence, an effective remedy for RILI is needed. Mesenchymal stromal cells (MSCs) are multipotent adult stem cells that have been demonstrated to be an effective treatment in some disease caused by tissue damage. However, unlike other injuries, RILI received limited therapeutic effects from implanted MSCs due to local hypoxia and extensive reactive oxygen species (ROS) in irradiated lungs. Since the poor survival of MSCs is primarily due to hypoxia and ROS generation, we hypothesize that persistent and adaptive hypoxia treatment induces enhanced resistance to hypoxic stress in implanted MSC. The aim of this study is to investigate whether persistent and adaptive hypoxia treatment of bmMSCs prior to their transplantation in injured mice enhanced survival and improved curative effects in RILI. Methods: Primary bmMSCs were obtained from the marrow of six-week-old male C57BL6/J mice and were cultured either under normoxic conditions (21% O-2) or hypoxic conditions (2.5% O-2). Mice were injected with normoxia/hypoxia MSCs after thoracic irradiation (20 Gy). The therapeutic effects of MSCs on RILI were assessed by pathological examinations that included H&E staining, Masson staining and alpha-SMA staining; meanwhile, B. Li, C. Li, M. Zhu and Y. Zhang contributed equally to this work. inflammatory factors were measured using an ELISA. The morphology of MSCs in vitro was recorded using a microscope and identified by flow cytometry, cell viability was measured using the CCK-8 assay, the potential for proliferation was detected by the EdU assay, and ROS levels were measured using a ROS fluorogenic probe. In addition, HIF-1 alpha and several survival pathway proteins (Akt, p-Akt, Caspase-3) were also detected by western blotting. Results: Implanted MSCs alleviated both early radiation-induced pneumonia and late pulmonary fibrosis. However, hypoxia MSCs displayed a more pronounced therapeutic effect compared to normoxia MSCs. Compared to normoxia MSCs, the hypoxia MSCs demonstrated greater cell viability, an enhanced proliferation potential, decreased ROS levels and increased resistance to hypoxia and ROS stress. In addition, hypoxia MSCs achieved higher activation levels of HIF-1 alpha and Akt, and HIF-1 alpha played a critical role in the development of resistance. Conclusion: Hypoxia enhances the therapeutic effect of mesenchymal stromal cells on radiation-induced lung injury by promoting MSC proliferation and improving their antioxidant ability, mediated by HIF-1 alpha. (c) 2017 The Author(s) Published by S. Karger AG, Basel