Background:: Acute kidney injury (AKI) is characterized by inflammatory infiltration and damage and death of renal tubular epithelial cells (RTECs), in which hypoxia plays an important role. Deferoxamine (DFO) is a well-accepted chemical hypoxia-mimetic agent. Mesenchymal stem cell-conditioned medium (MSC-CM) can reduce local inflammation and repair tissue. In this study, we explored the effect and molecular mechanism of MSC-CM-mediated protection of RTECs under DFO-induced hypoxia. background: Acute kidney injury (AKI) is characterized by inflammatory infiltration, and damage and death of renal tubular epithelial cells (RTECs), in which hypoxia plays an important role. Deferoxamine (DFO)-induced hypoxia injury is a well-accepted renal injury model in vitro. Mesenchymal stem cell - conditioned medium (MSC-CM) can reduce local inflammation and repair tissue. Methods:: Rat renal proximal tubule NRK-52E cells were treated with different concentrations of DFO for 24 hours, followed by evaluation of RTEC injury, using a Cell Counting Kit-8 (CCK-8) to detect cell viability and western blotting to evaluate the expression of transforming growth factor- beta 1 (TGF-β1), α-smooth muscle actin (α-SMA), and hypoxia-inducible factor-1 alpha (HIF-1α) in NRK-52E cells. Then, three groups of NRK-52E cells were used in experiments, including normal control (NC), 25 μM DFO, and 25 μM DFO + MSC-CM. MSC-CM was obtained from the human umbilical cord. MSC-CM was used to culture cells for 12 hours before DFO treatment, then fresh MSC-CM and 25 μM DFO were added, and cells were cultured for another 24 hours before analysis. Results:: Western blotting and cellular immunofluorescence staining showed culture of NRK-52E cells in 25 μM DFO for 24 hours induced HIF-1α and nuclear receptor coactivator-1 (NCoA-1), simulating hypoxia. MSC-CM could inhibit the DFO-induced up-regulation of α-SMA, TGF-β1, HIF-1α and NCoA-1. method: Rat renal proximal tubule NRK-52E cells were treated with different concentrations of DFO for 24 hours, followed by evaluation of RTEC injury, using a Cell Counting Kit-8 (CCK-8) to detect cell viability, and western blotting to evaluate the expression of transforming growth factor-beta 1 (TGF-β1), α-smooth muscle actin (α-SMA), and hypoxia-inducible factor-1 alpha (HIF-1α) in NRK-52E cells. Then three groups of NRK-52E cells were used in experiments, including normal control (NC), 25 μM DFO, and 25 μM DFO + MSC-CM. MSC-CM was obtained from human umbilical cord. MSC-CM was used to culture cells for 12 hours before DFO treatment, then fresh MSC-CM and 25 μM DFO were added and cells were cultured for another 24 hours before analysis. Conclusion:: Our results suggest that MSC-CM has a protective effect on RTECs by down-regulating HIF-1α and NCoA-1, which may be the harmful factors in renal injury. other: -

中文翻译：

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间充质干细胞条件培养基通过抑制缺氧诱导因子-1α和核受体辅激活因子-1保护肾小管上皮细胞

背景：急性肾损伤（AKI）以炎症浸润和肾小管上皮细胞（RTEC）损伤和死亡为特征，其中缺氧起着重要作用。去铁胺（DFO）是一种广为接受的化学拟缺氧剂。间充质干细胞条件培养基（MSC-CM）可以减轻局部炎症并修复组织。在本研究中，我们探讨了 MSC-CM 介导的 DFO 缺氧条件下对 RTEC 的保护作用和分子机制。背景：急性肾损伤（AKI）以炎症浸润、肾小管上皮细胞（RTEC）损伤和死亡为特征，其中缺氧起着重要作用。去铁胺（DFO）引起的缺氧损伤是一种公认​​的体外肾损伤模型。间充质干细胞-条件培养基（MSC-CM）可以减轻局部炎症并修复组织。方法：：大鼠肾近曲小管NRK-52E细胞用不同浓度的DFO处理24小时，然后评估RTEC损伤，使用细胞计数试剂盒8（CCK-8）检测细胞活力，并用免疫印迹法评估RTEC损伤情况。 NRK-52E 细胞中转化生长因子 - β 1 (TGF-β1)、α-平滑肌肌动蛋白 (α-SMA) 和缺氧诱导因子 1 α (HIF-1α) 的表达。然后，使用三组NRK-52E细胞进行实验，包括正常对照（NC）、25μM DFO和25μM DFO + MSC-CM。MSC-CM 是从人脐带中获得的。DFO处理前用MSC-CM培养细胞12小时，然后加入新鲜MSC-CM和25μM DFO，再培养细胞24小时后进行分析。结果：Western blotting 和细胞免疫荧光染色显示，NRK-52E 细胞在 25 μM DFO 中培养 24 小时诱导 HIF-1α 和核受体辅激活剂-1 (NCoA-1)，模拟缺氧。MSC-CM 可以抑制 DFO 诱导的 α-SMA、TGF-β1、HIF-1α 和 NCoA-1 的上调。方法：用不同浓度的DFO处理大鼠肾近曲小管NRK-52E细胞24小时，评估RTEC损伤情况，使用细胞计数试剂盒（CCK-8）检测细胞活力，免疫印迹法评估RTEC损伤情况NRK-52E 细胞中转化生长因子-β 1 (TGF-β1)、α-平滑肌肌动蛋白 (α-SMA) 和缺氧诱导因子 1 α (HIF-1α) 的表达。然后使用三组NRK-52E细胞进行实验，包括正常对照（NC）、25μM DFO和25μM DFO+MSC-CM。MSC-CM 是从人脐带中获得的。DFO处理前使用MSC-CM培养细胞12小时，然后加入新鲜MSC-CM和25μM DFO，再培养细胞24小时，然后进行分析。结论::我们的结果表明MSC-CM通过下调HIF-1α和NCoA-1对RTECs具有保护作用，而HIF-1α和NCoA-1可能是肾损伤的有害因素。其他： -