Department of Burn Cosmetology, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, 264000, China.
Wound Repair Regen. 2020 Jan;28(1):39-48. doi: 10.1111/wrr.12765. Epub 2019 Nov 12.
To study the effects of mir-27b on angiogenesis and fibroblast activation and to explore its further mechanism. Humanmicrovascular endothelial cell (HMEC)-1 and humannormal skin fibroblast (BJ) cells were treated with mir-27b inhibitor negative control reagent, mir-27b inhibitor, LY294002, and mir-27b inhibitor + LY294002, respectively. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used to detect the T-cell proliferation. The migration ability was detected by Scratch assays. The angiogenesis of HMEC-1 cells was observed by in vitro tube formation assay. The mRNA and protein expression of vascular endothelial growth factor (VEGF) in HMEC-1 cells and the mRNA and protein expression of collagen I, collagen III, α-SMA, and MMP1 in BJ cells were detected by quantitativereal-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Meanwhile, the PI3K/protein kinase B (AKT) pathway-related proteins were also detected by Western blot. The proliferation, migration, angiogenesis, the mRNA and protein expression of VEGF and the protein expression of p-PI3K and p-AKT in HMEC-1 cells were increased after treated with mir-27b inhibitor. Meanwhile, the proliferation, migration, and the protein expression of collagen I, collagen III, α-SMA, MMP1, p-PI3K, and p-AKT in BJ cells were increased after treated with mir-27b inhibitor. However, the angiogenesis and fibroblast activation of mir-27b inhibitor was reversed by LY294002, and the activate effect to PI3K/AKT pathway was also inhibited. Down-regulation of mir-27b could promote angiogenesis and fibroblast activation, and its mechanism is related to activate PI3K/AKT signaling pathway.
为了研究 mir-27b 对血管生成和成纤维细胞激活的影响，并进一步探讨其机制。分别用 mir-27b 抑制剂阴性对照试剂、mir-27b 抑制剂、LY294002 处理人微血管内皮细胞（HMEC-1）和正常人皮肤成纤维细胞（BJ），用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测 T 细胞增殖，划痕实验检测迁移能力，体外管形成实验观察 HMEC-1 细胞的血管生成。通过定量实时聚合酶链反应（qRT-PCR）和 Western blot 检测 HMEC-1 细胞中血管内皮生长因子（VEGF）的 mRNA 和蛋白表达，BJ 细胞中胶原 I、胶原 III、α-SMA 和 MMP1 的 mRNA 和蛋白表达，同时通过 Western blot 检测 PI3K/蛋白激酶 B（AKT）通路相关蛋白。mir-27b 抑制剂处理后，HMEC-1 细胞增殖、迁移、血管生成、VEGF 的 mRNA 和蛋白表达增加，同时 BJ 细胞增殖、迁移增加，胶原 I、胶原 III、α-SMA、MMP1、p-PI3K 和 p-AKT 蛋白表达增加。然而，LY294002 逆转了 mir-27b 抑制剂的血管生成和成纤维细胞激活作用，并抑制了 PI3K/AKT 通路的激活。下调 mir-27b 可促进血管生成和成纤维细胞激活，其机制与激活 PI3K/AKT 信号通路有关。
