Department of Bone Surgery, The First Hospital of Harbin City, Harbin, Heilongjiang 150010, P.R. China.
Department of Plastic Surgery, The First Hospital of Harbin City, Harbin, Heilongjiang 150010, P.R. China.
Mol Med Rep. 2017 Dec;16(6):8826-8832. doi: 10.3892/mmr.2017.7701. Epub 2017 Oct 3.
A variety of pulsed electromagnetic fields (PEMFs) have been experimentally and clinically used in an effort to promote wound healing, although the mechanisms involved remain unknown. The aim of the present study was to investigate the action of a novel protocol of co‑treatment with PEMFs and hydroxytyrosol (HTY) on the proliferation and differentiation potential of human umbilical vein endothelial cells (HUVECs). The HUVECs were assigned randomly into three groups: Control, PEMF‑treated and PEMF + HT‑treated. The intensity of the electromagnetic field used in this protocol was 2.25 mT, the frequency of the bursts was 50 Hz and the application time was 15 min. A Cell Counting kit‑8 (CCK‑8) assay was used to assess cell proliferation, and cell apoptosis was analyzed by TUNEL apoptosis assay kit and calcein‑acetoxymethyl/propidium iodide dual‑staining assay. In addition, protein and mRNA expression levels of protein kinase B (Akt), mechanistic target of rapamycin (mTOR), transforming growth factor (TGF)‑β1 and p53 were determined by western blotting and reverse transcription‑quantitative polymerase chain reaction assays, respectively. The CCK‑8 assay demonstrated that HTY contributed to HUVEC proliferation mediated by PEMFs in a time‑dependent manner. The Transwell assay and scratch wound results demonstrated that co‑treatment of HTY and PEMFs could increase HUVEC migration. Furthermore, the levels of apoptotic cells were reversed by pre‑incubation with HTY in the PEMF treatment group, while PEMF treatment alone had no such effect. The proteins and mRNA expression levels of Akt, mTOR, TGF‑β1 were elevated in co‑treatment of HTY and PEMFs, whereas there was no effect on levels of p53. Therefore, the results indicated that combined exposure of HUVECs to PEMFs and HTY exerted protective effects in HUVECs by promoting cell proliferation and inhibiting apoptosis. In conclusion, to the best of our knowledge, the present study was the first to demonstrate the beneficial roles of HTY and PEMF combined treatment in HUVECs, which may represent an effective treatment for wound healing.
各种脉冲电磁场 (PEMFs) 已在实验和临床中被用于促进伤口愈合，尽管其涉及的机制仍不清楚。本研究的目的是研究新型 PEMFs 与羟基酪醇 (HTY) 联合处理方案对人脐静脉内皮细胞 (HUVEC) 增殖和分化潜力的作用。将 HUVEC 随机分为三组：对照组、PEMF 处理组和 PEMF+HT 处理组。本方案中使用的电磁场强度为 2.25 mT，爆发频率为 50 Hz，应用时间为 15 分钟。使用细胞计数试剂盒 -8 (CCK-8) 测定法评估细胞增殖，通过 TUNEL 凋亡测定试剂盒和 calcein-acetoxymethyl/propidium iodide 双重染色测定法分析细胞凋亡。此外，通过 Western blot 和逆转录定量聚合酶链反应 (qPCR) 测定法分别测定蛋白激酶 B (Akt)、雷帕霉素靶蛋白 (mTOR)、转化生长因子 (TGF)-β1 和 p53 的蛋白和 mRNA 表达水平。CCK-8 测定法表明，HTY 以时间依赖性方式促进 PEMFs 介导的 HUVEC 增殖。Transwell 测定法和划痕伤口结果表明，HTY 和 PEMFs 的联合处理可增加 HUVEC 迁移。此外，在用 HTY 预处理 PEMF 处理组中，可逆转细胞凋亡水平，而单独使用 PEMF 处理则没有这种作用。在 HTY 和 PEMFs 的联合处理中，Akt、mTOR、TGF-β1 的蛋白和 mRNA 表达水平升高，而 p53 的水平没有影响。因此，结果表明，HUVEC 暴露于 PEMFs 和 HTY 的联合作用可通过促进细胞增殖和抑制凋亡对 HUVEC 发挥保护作用。总之，据我们所知，本研究首次证明了 HTY 和 PEMF 联合治疗在 HUVEC 中的有益作用，这可能代表了一种有效的伤口愈合治疗方法。
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