Department of Endocrinology and Metabolism, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.
School of Pharmaceutical Sciences of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.
Int J Mol Med. 2019 May;43(5):2187-2198. doi: 10.3892/ijmm.2019.4140. Epub 2019 Mar 20.
Metformin serves an important role in improving the functions of endothelial progenitor cells (EPCs). MicroRNAs (miRNAs), small non‑coding RNAs, have been investigated as significant regulators of EPC vascular functions. The present study investigated the molecular crosstalk between metformin and miRNA‑130a (miR‑130a) in the functions of EPCs exposed to palmitic acid (PA). Isolated EPCs were treated with metformin, PA, and metformin + PA, respectively. Cell Counting Kit‑8, Transwell and Matrigel assays were performed to detect the proliferation, migration and tube formation ability of EPCs following different treatments. The expression of miR‑130a, phosphatase and tensin homolog (PTEN) and phosphorylated‑AKT was analyzed by reverse transcription‑quantitative polymerase chain reaction and western blotting. The specific mechanism underlying the function of metformin in EPCs was further elucidated by transfecting miR‑130a mimics and inhibitor to overexpress and inhibit the expression of miR‑130a in EPCs, respectively. EPCs exhibited impaired functions of proliferation (P<0.01 compared with the control), migration (P<0.01 compared with the control) and tube formation (P<0.01 compared with the control) following treatment with PA, and the expression levels of miR‑130a and PTEN were decreased and increased, respectively. However, the presence of metformin, or the overexpression of miR‑130a using miR‑130a mimic alleviated the impairment of angiogenesis and proliferation, decreased the expression of PTEN and activated the phosphoinositide‑3 kinase/AKT pathway in EPCs exposed to PA. By contrast, downregulating the expression of miR‑130a with a miR‑130a inhibitor reversed the metformin‑mediated protection. These results demonstrate the beneficial effect of miR‑130a/PTEN on EPC functions, which can be regulated by metformin. The effects of metformin on improving PA‑induced EPC dysfunction are mediated by miR‑130a and PTEN, which may assist in the prevention and/or treatment of diabetic vascular disease.
二甲双胍在改善内皮祖细胞 (EPC) 功能方面发挥着重要作用。微小 RNA (miRNA) 是一种小的非编码 RNA，已被研究为调节 EPC 血管功能的重要调控因子。本研究探讨了二甲双胍和 miRNA-130a（miR-130a）在暴露于棕榈酸 (PA) 的 EPC 功能中的分子相互作用。分别用二甲双胍、PA 和二甲双胍+PA 处理分离的 EPC。通过细胞计数试剂盒-8、Transwell 和 Matrigel 测定法检测不同处理后 EPC 的增殖、迁移和管形成能力。采用逆转录-定量聚合酶链反应和蛋白质印迹法分析 miR-130a、磷酸酶和张力蛋白同源物 (PTEN) 和磷酸化-AKT 的表达。通过转染 miR-130a 模拟物和抑制剂分别过表达和抑制 EPC 中 miR-130a 的表达，进一步阐明了二甲双胍在 EPC 中的作用的具体机制。用 PA 处理后，EPC 的增殖功能受损（与对照组相比，P<0.01）、迁移功能受损（与对照组相比，P<0.01）和管形成功能受损（与对照组相比，P<0.01），miR-130a 和 PTEN 的表达水平分别降低和升高。然而，二甲双胍的存在，或使用 miR-130a 模拟物过表达 miR-130a 可减轻 EPC 暴露于 PA 后血管生成和增殖受损，降低 PTEN 的表达并激活磷酸肌醇-3 激酶/AKT 通路。相反，用 miR-130a 抑制剂下调 miR-130a 的表达可逆转二甲双胍介导的保护作用。这些结果表明 miR-130a/PTEN 对 EPC 功能具有有益作用，而这种作用可被二甲双胍调节。二甲双胍改善 PA 诱导的 EPC 功能障碍的作用是通过 miR-130a 和 PTEN 介导的，这可能有助于预防和/或治疗糖尿病血管疾病。
