Wu Shenhao, Luo Xiaomei, Liu Yanwen, Gao Jing
The Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi, 830000, China.
Xinjiang Medical University, Urumqi, 830000, China.
Ir J Med Sci. 2025 Sep 3. doi: 10.1007/s11845-025-04077-1.
Diabetic vascular complications present significant clinical challenges, including limited treatment efficacy, high postoperative restenosis rates, and delayed early diagnosis. This study investigates CXCR4-modified adipose-derived mesenchymal stem cells (AMSCs/CXCR4) in regulating pathological endothelial proliferation under hyperglycemic conditions.
The purpose is to provide new mechanism insights and potential therapeutic targets for early intervention of diabetes-related vascular diseases.
The CXCR4-overexpressing plasmid was generated via XhoI/EcoRI double digestion, T4 ligation, and column purification, then transfected into AMSCs using Lipofectamine® 3000 to enhance exosome secretion. These exosomes were co-cultured with HG-treated HUVECs. Cell viability and apoptosis were assessed by CCK8 and flow cytometry. AKT/mTOR pathway proteins (total/phosphorylated) were analyzed via Western blot, while qRT-PCR quantified miRNA320, VEGF, and IGF-1 expression.
Chronic high glucose stimulated abnormal endothelial cell proliferation (CCK-8/flow cytometry), which was suppressed by AMSCs/CXCR4, reducing proliferation and elevating apoptosis ( 21.723 ± 1.061% apoptosis rate)). High glucose downregulated miRNA320, but AMSCs/CXCR4 restored its expression ( 0.937 ± 0.056 vs. other groups, P < 0.05). Increased miRNA320 correlated with reduced VEGF ((1.101 ± 0.142) and IGF-1 (1.074 ± 0.084) levels, confirming miRNA320-mediated inhibition. Notably, activation of the AKT/mTOR pathway proteins was not affected, indicating that AMSCs/CXCR4 directly inhibited the activity of VEGF and IGF-1 in HUVECs via miRNA320.
CXCR4 boosts exosome release from AMSCs. Although AMSCs/CXCR4 did not alter AKT/mTOR signaling, their miRNA320-loaded exosomes blocked IGF-1/VEGF activity. This study uncovers a CXCR4-miRNA320 axis in diabetic vascular dysfunction, highlighting exosome-based therapy and miRNA320 as a targeted strategy for vascular complications.
糖尿病血管并发症带来了重大临床挑战，包括治疗效果有限、术后再狭窄率高以及早期诊断延迟。本研究调查了CXCR4修饰的脂肪来源间充质干细胞（AMSCs/CXCR4）在高血糖条件下对病理性内皮细胞增殖的调节作用。
旨在为糖尿病相关血管疾病的早期干预提供新的机制见解和潜在治疗靶点。
通过XhoI/EcoRI双酶切、T4连接和柱纯化生成CXCR4过表达质粒，然后使用Lipofectamine® 3000将其转染到AMSCs中以增强外泌体分泌。将这些外泌体与高糖处理的人脐静脉内皮细胞（HUVECs）共培养。通过CCK8和流式细胞术评估细胞活力和凋亡。通过蛋白质印迹法分析AKT/mTOR通路蛋白（总/磷酸化），而qRT-PCR定量miRNA320、血管内皮生长因子（VEGF）和胰岛素样生长因子-1（IGF-1）的表达。
慢性高糖刺激内皮细胞异常增殖（CCK-8/流式细胞术），AMSCs/CXCR4可抑制这种增殖，减少增殖并提高凋亡率（凋亡率为21.723±1.061%）。高糖下调miRNA320，但AMSCs/CXCR4可恢复其表达（与其他组相比为0.937±0.056，P<0.05）。miRNA320增加与VEGF（(1.101±0.142)）和IGF-1（1.074±0.084）水平降低相关，证实了miRNA320介导的抑制作用。值得注意的是，AKT/mTOR通路蛋白的激活未受影响，表明AMSCs/CXCR4通过miRNA320直接抑制HUVECs中VEGF和IGF-1的活性。
CXCR4促进AMSCs释放外泌体。虽然AMSCs/CXCR4未改变AKT/mTOR信号传导，但其负载miRNA320的外泌体阻断了IGF-1/VEGF活性。本研究揭示了糖尿病血管功能障碍中的CXCR4-miRNA320轴，突出了基于外泌体的治疗以及miRNA320作为血管并发症的靶向策略。
