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HIF1α overexpression enhances diabetic wound closure in high glucose and low oxygen conditions by promoting adipose-derived stem cell paracrine function and survival.
Stem Cell Research & Therapy ( IF 7.5 ) Pub Date : 2020-04-05 , DOI: 10.1186/s13287-020-01654-2
Jin Xu 1 , Xiaoyu Liu 2 , Feng Zhao 3 , Ying Zhang 4 , Zhe Wang 4
Affiliation  

Adipose-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. Adipose 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. In 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. HIF1α 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.

中文翻译:

HIF1α 过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。

由于干细胞的旁分泌功能,脂肪源性干细胞 (ADSC) 移植是促进伤口愈合的有前景的策略。然而,葡萄糖对干细胞旁分泌功能和存活的影响会导致糖尿病患者的伤口闭合受损,从而限制了 ADSC 移植的疗效。缺氧诱导因子(HIF)1α在伤口愈合中起重要作用,在本研究中,我们研究了高糖低氧条件下HIF1α过表达对ADSCs的影响。脂肪样品取自 BALB/C 小鼠,通过消化体外培养 ADSC。通过转导诱导对照和过表达 HIF1α 的 ADSC。通过定量逆转录聚合酶链反应和蛋白质印迹分析在高葡萄糖和低氧条件下对照和过表达HIF1α的ADSC中血管生成生长因子的mRNA和蛋白质水平。使用体外共培养模型评估高糖条件下 ADSC HIF1α 过表达对小鼠主动脉内皮细胞 (MAECs) 增殖和迁移的影响。分别使用 2,7-二氯二氢荧光素二乙酸酯染色和酶联免疫吸附测定法观察 ADSC 中的细胞内活性氧 (ROS) 和 8-羟基脱氧鸟苷 (8-OHdG) 水平。通过流式细胞术进行细胞凋亡和细胞周期分析测定。使用体内全层皮肤缺损小鼠模型来评估移植的 ADSCs 对糖尿病伤口闭合的影响。在体外,ADSCs 中 HIF1α 过表达显着增加了高糖抑制的血管内皮生长因子 A、成纤维细胞生长因子 2 和 CXC 基序趋化因子配体 12 的表达。ADSCs 中的 HIF1α 过表达减轻了高糖诱导的 MAEC 增殖和迁移缺陷,并显着抑制了 ADSC ROS 和 8-OHdG 水平,从而减少细胞凋亡并提高存活率。在体内,移植前 ADSC 中的 HIF1α 过表达显着增强了血管生成生长因子的表达,促进了糖尿病小鼠的伤口闭合。ADSCs 中 HIF1α 过表达可有效缓解高糖诱导的旁分泌功能障碍,减少氧化应激和随后的 DNA 损伤,提高 ADSCs 的活力,并增强 ADSCs 对糖尿病伤口愈合的治疗效果。ADSCs 中 HIF1α 过表达显着增加了高糖抑制的血管内皮生长因子 A、成纤维细胞生长因子 2 和 CXC 基序趋化因子配体 12 的表达。ADSCs 中的 HIF1α 过表达减轻了高糖诱导的 MAEC 增殖和迁移缺陷,并显着抑制了 ADSC ROS 和 8-OHdG 水平,从而减少细胞凋亡并提高存活率。在体内,移植前 ADSC 中的 HIF1α 过表达显着增强了血管生成生长因子的表达,促进了糖尿病小鼠的伤口闭合。ADSCs 中 HIF1α 过表达可有效缓解高糖诱导的旁分泌功能障碍,减少氧化应激和随后的 DNA 损伤,提高 ADSCs 的活力,并增强 ADSCs 对糖尿病伤口愈合的治疗效果。ADSCs 中 HIF1α 过表达显着增加了高糖抑制的血管内皮生长因子 A、成纤维细胞生长因子 2 和 CXC 基序趋化因子配体 12 的表达。ADSCs 中的 HIF1α 过表达减轻了高糖诱导的 MAEC 增殖和迁移缺陷,并显着抑制了 ADSC ROS 和 8-OHdG 水平,从而减少细胞凋亡并提高存活率。在体内,移植前 ADSC 中的 HIF1α 过表达显着增强了血管生成生长因子的表达,促进了糖尿病小鼠的伤口闭合。ADSCs 中 HIF1α 过表达可有效缓解高糖诱导的旁分泌功能障碍,减少氧化应激和随后的 DNA 损伤,提高 ADSCs 的活力,并增强 ADSCs 对糖尿病伤口愈合的治疗效果。
更新日期:2020-04-22
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