HIF1α overexpression enhances diabetic wound closure in high glucose and low oxygen conditions by promoting adipose-derived stem cell paracrine function and survival
HIF1α overexpression enhances diabetic wound closure in high glucose and low oxygen conditions by promoting adipose-derived stem cell paracrine function and survival
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HIF1α 过度表达通过促进脂肪干细胞旁分泌功能和存活来增强高葡萄糖和低氧条件下的糖尿病伤口闭合
DOI：
10.1186/s13287-020-01654-2
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发表时间：
2020-04-05
影响因子：
7.5
通讯作者：
Wang, Zhe
中科院分区：
文献类型：
--
作者：
Xu, Jin;Liu, Xiaoyu;Wang, Zhe
背景脂肪干细胞（adipose-derived stem cells，ADSC）移植由于其旁分泌功能而成为促进创伤愈合的一种有效方法。然而，葡萄糖对干细胞旁分泌功能和生存的相关影响导致糖尿病患者的伤口闭合受损，从而限制了ADSC移植的功效。低氧诱导因子（Hypoxia-inducible factor，HIF）1α在创伤愈合过程中起重要作用，本研究探讨高糖低氧条件下HIF 1 α过表达对脂肪干细胞的影响。通过转导诱导对照和HIF 1 α过表达的ADSC。采用定量逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹法（Western blotting）分析高糖低氧条件下对照组和HIF 1 α过表达组ADSCs中血管生成因子的mRNA和蛋白水平。采用体外共培养模型，观察高糖环境下ADSC HIF 1 α过表达对小鼠主动脉内皮细胞（MAECs）增殖和迁移的影响。分别采用2，7-二氯二氢荧光素二乙酸酯染色法和酶联免疫吸附法检测ADSC细胞内活性氧（ROS）和8-羟基脱氧鸟苷（8-OHdG）水平。流式细胞仪检测细胞凋亡和细胞周期。采用全层皮肤缺损小鼠模型，观察移植脂肪干细胞对糖尿病创面愈合的影响。结果在体外，HIF 1 α过表达的脂肪干细胞可显著增加血管内皮生长因子A、成纤维细胞生长因子2和C-X-C基序趋化因子配体12的表达，而高糖可抑制这些因子的表达。在ADSC中HIF 1 α过表达可减轻高糖诱导的MAEC增殖和迁移缺陷，并显著抑制ADSC ROS和8-OHdG水平，从而减少凋亡并提高存活率。结论HIF 1 α过表达的ADSCs能有效缓解高糖诱导的旁分泌功能障碍，减轻氧化应激和DNA损伤，提高ADSCs的存活率，增强ADSCs对糖尿病创面愈合的治疗作用。
BackgroundAdipose-derived stem cell (ADSC) transplantation is a promising strategy to promote wound healing because of the paracrine function of stem cells. However, glucose-associated effects on stem cell paracrine function and survival contribute to impaired wound closure in patients with diabetes, limiting the efficacy of ADSC transplantation. Hypoxia-inducible factor (HIF)1α plays important roles in wound healing, and in this study, we investigated the effects of HIF1α overexpression on ADSCs in high glucose and low oxygen conditions.MethodsAdipose samples were obtained from BALB/C mice, and ADSCs were cultured in vitro by digestion. Control and HIF1α-overexpressing ADSCs were induced by transduction. The mRNA and protein levels of angiogenic growth factors in control and HIF1α-overexpressing ADSCs under high glucose and low oxygen conditions were analyzed by quantitative reverse transcription-polymerase chain reaction and western blotting. The effects of ADSC HIF1α overexpression on the proliferation and migration of mouse aortic endothelial cells (MAECs) under high glucose were evaluated using an in vitro coculture model. Intracellular reactive oxygen species (ROS) and 8-hydroxydeoxyguanosine (8-OHdG) levels in ADSCs were observed using 2,7-dichlorodihydrofluorescein diacetate staining and enzyme-linked immunosorbent assays, respectively. Apoptosis and cell cycle analysis assays were performed by flow cytometry. An in vivo full-thickness skin defect mouse model was used to evaluate the effects of transplanted ADSCs on diabetic wound closure.ResultsIn vitro, HIF1α overexpression in ADSCs significantly increased the expression of vascular endothelial growth factor A, fibroblast growth factor 2, and C-X-C motif chemokine ligand 12, which were inhibited by high glucose. HIF1α overexpression in ADSCs alleviated high glucose-induced defects in MAEC proliferation and migration and significantly suppressed ADSC ROS and 8-OHdG levels, thereby decreasing apoptosis and enhancing survival. In vivo, HIF1α overexpression in ADSCs prior to transplantation significantly enhanced angiogenic growth factor expression, promoting wound closure in diabetic mice.ConclusionsHIF1α overexpression in ADSCs efficiently alleviates high glucose-induced paracrine dysfunction, decreases oxidative stress and subsequent DNA damage, improves viability, and enhances the therapeutic effects of ADSCs on diabetic wound healing.