AMSC/CXCR4-derived exosomes and miRNA-320 regulate pathological angiogenesis in diabetes.

BACKGROUND

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.

AIMS

The purpose is to provide new mechanism insights and potential therapeutic targets for early intervention of diabetes-related vascular diseases.

METHODS

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.

RESULTS

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.

CONCLUSIONS

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.