Department of Nephrology, Second Hospital of Lanzhou University, Lanzhou 730030, Gansu, China.
Curr Gene Ther. 2024;24(2):159-177. doi: 10.2174/0115665232258527230919071328.
Diabetic nephropathy (DN) is one of the microvascular complications of diabetes. Endothelial-mesenchymal transition (EndMT) and endothelial damage lead to abnormal angiogenesis in DN.
This study aimed to investigate the role of exosome miR-30a-5p in high glucose (HG)-induced glomerular endothelial cells (GECs) dysfunction and explore the underlying mechanisms.
GECs were cultured in normal glucose (5.5 mM) and HG (30 mM) conditions. The recipient GECs were transfected with exosome or miR-30a-5p mimic/inhibitor and then detected by using CCK-8 and flow cytometry assay. Luciferase analysis was used to verify miR-30a-5p acted on notch homolog protein 1 (Notch1). RT-qPCR and Western blot were used to detect the expression of VE-cadherin, α-SMA, vascular endothelial growth factor (VEGF) and Notch1. In vivo, exosome miR-30a-5p was administered to DN mice, and periodic acid-Schiff (PAS) staining, UTP levels, and HbA1c levels were measured.
The expression of miR-30a-5p was downregulated in HG-treated GECs. Exosome miR-30a-5p significantly promoted cell proliferation, and migration and reduced apoptosis of GECs under HG conditions. MiR-30a-5p directly targeted the 3-UTR region of Notch1. Exosome miR-30a-5p reduced the expression levels of Notch1 and VEGF, both at mRNA and protein levels. Furthermore, exosome miR-30a-5p inhibited HG-induced EndMT, as evidenced by increased VE-cadherin and reduced α-SMA. In vivo studies demonstrated that exosome miR-30a-5p reduced serum HbA1c levels and 24-hour urine protein quantification.
This study provides evidence that exosome miR-30a-5p suppresses EndMT and abnormal angiogenesis of GECs by modulating the Notch1/VEGF signaling pathway. These findings suggest that exosome miR-30a-5p could be a potential therapeutic strategy for the treatment of DN.
糖尿病肾病（DN）是糖尿病的微血管并发症之一。内皮-间充质转化（EndMT）和内皮损伤导致 DN 中异常血管生成。
本研究旨在探讨外泌体 miR-30a-5p 在高糖（HG）诱导的肾小球内皮细胞（GEC）功能障碍中的作用，并探讨其潜在机制。
将 GEC 在正常葡萄糖（5.5mM）和 HG（30mM）条件下培养。用外泌体或 miR-30a-5p 模拟物/抑制剂转染受体 GEC，然后通过 CCK-8 和流式细胞术检测。荧光素酶分析用于验证 miR-30a-5p 作用于 Notch 同源蛋白 1（Notch1）。RT-qPCR 和 Western blot 用于检测 VE-钙黏蛋白、α-SMA、血管内皮生长因子（VEGF）和 Notch1 的表达。在体内，给予 DN 小鼠外泌体 miR-30a-5p，测量过碘酸-Schiff（PAS）染色、UTP 水平和 HbA1c 水平。
HG 处理的 GEC 中 miR-30a-5p 的表达下调。外泌体 miR-30a-5p 显著促进 HG 条件下 GEC 的增殖、迁移和减少凋亡。miR-30a-5p 可直接靶向 Notch1 的 3'-UTR 区域。外泌体 miR-30a-5p 降低 Notch1 和 VEGF 的 mRNA 和蛋白水平。此外，外泌体 miR-30a-5p 抑制 HG 诱导的 EndMT，表现为 VE-钙黏蛋白增加和 α-SMA 减少。体内研究表明，外泌体 miR-30a-5p 降低了血清 HbA1c 水平和 24 小时尿蛋白定量。
本研究提供的证据表明，外泌体 miR-30a-5p 通过调节 Notch1/VEGF 信号通路抑制 GEC 的 EndMT 和异常血管生成。这些发现表明，外泌体 miR-30a-5p 可能成为治疗 DN 的潜在治疗策略。
