Department of Basic Medicine, Jiangsu College of Nursing, Huai'an, Jiangsu 223005, P.R. China.
Mol Med Rep. 2024 Dec;30(6). doi: 10.3892/mmr.2024.13347. Epub 2024 Oct 4.
The present study aimed to investigate the role of microRNA (miR)‑221‑3p in endothelial progenitor cells (EPCs) treated with lipoprotein(a) [LP(a)]. EPCs were identified using immunofluorescence assays and miR‑221‑3p levels were measured using reverse transcription‑quantitative PCR. EPC migration was detected using Transwell assays, proliferation was measured by staining with 5‑ethynyl‑2'‑deoxyuridine and adhesion was assessed by microscopy. Flow cytometry was used to measure apoptosis and protein expression was detected using western blotting. A dual‑luciferase reporter assay was used to confirm the target interactions. The proliferation, migration, adhesion and angiogenesis of EPCs were decreased, and apoptosis was increased after treatment with LP(a). These effects were weakened by transfection with miR‑221‑3p inhibitor. The negative effects of LP(a) on EPCs were also weakened by overexpression of silent information regulator 1 (SIRT1). Inhibition of the RAF/MEK/ERK signaling pathway blocked the effects of SIRT1 overexpression. In conclusion, miR‑221‑3p inhibitor transfection activated the RAF/MEK/ERK signaling pathway through SIRT1, promoted the proliferation, migration, adhesion and angiogenesis of EPCs, and reduced apoptosis.
本研究旨在探讨脂蛋白(a) [LP(a)]处理的内皮祖细胞(EPCs)中微小 RNA (miR) -221-3p 的作用。通过免疫荧光法鉴定 EPCs,采用逆转录 - 定量 PCR 测定 miR-221-3p 水平。采用 Transwell 检测 EPC 迁移,用 5-乙炔基-2'-脱氧尿苷染色法检测增殖,用显微镜评估黏附。采用流式细胞术检测细胞凋亡,采用 Western blot 检测蛋白表达。采用双荧光素酶报告基因检测证实靶基因相互作用。LP(a) 处理后 EPCs 的增殖、迁移、黏附和血管生成减少,细胞凋亡增加。miR-221-3p 抑制剂转染可减弱这些作用。沉默信息调节因子 1 (SIRT1) 的过表达也减弱了 LP(a) 对 EPCs 的负作用。RAF/MEK/ERK 信号通路的抑制阻断了 SIRT1 过表达的作用。综上所述,miR-221-3p 抑制剂转染通过 SIRT1 激活 RAF/MEK/ERK 信号通路,促进 EPCs 的增殖、迁移、黏附和血管生成,减少凋亡。