AUTHOR=Li Yaodong , Chen Chuanzhen , Kong Jingbo , Huang Junjie , Liu Liguo , Liu Cunfa , Dai Bing , Huang Mei 

TITLE=Bone marrow mesenchymal stem cell-derived exosomal microRNA-335 alleviates vascular calcification by targeting SP1

JOURNAL=Frontiers in Cardiovascular Medicine

VOLUME=Volume 12 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/cardiovascular-medicine/articles/10.3389/fcvm.2025.1572045

DOI=10.3389/fcvm.2025.1572045

ISSN=2297-055X

ABSTRACT=BackgroundVascular calcification (VC) is a critical pathological characteristic of cardiovascular diseases like atherosclerosis, frequently linked to phenotypic alterations in vascular smooth muscle cells (VSMCs) and the activation of bone-forming genes. Exosomes derived from bone marrow mesenchymal stem cells (BMSCs) have been shown to significantly attenuate VSMC calcification.MethodsTo investigate whether BMSC-derived exosomes mitigate VSMC calcification through microRNAs (miRNAs) regulation, we developed an in vitro model using β-glycerophosphate-induced calcification and an in vivo model using vitamin D3-induced calcification. Exosomes were extracted from BMSC culture media via ultracentrifugation and analyzed using transmission electron microscopy and particle size distribution assays.ResultsFunctional and phenotypic assessments revealed that BMSC-derived exosomes markedly reduced VSMC calcification. RT-qPCR analysis further indicated that BMSC-derived exosomes regulate VSMC calcification by modulating rno-miR-335 (miR-335). The miR-335 mimic notably suppressed the expression of the osteogenic regulator RUNX2 in VSMCs. Dual-luciferase reporter assays demonstrated that SP1 is a direct target of miR-335. Exosomal miR-335 inhibited SP1 expression, resulting in reduced mRNA and protein levels of RUNX2. In vivo studies confirmed that agomiR-335 treatment significantly lowered SP1 levels in the aorta of male SD rats, alleviating vitamin D3-induced VC.ConclusionThis study highlights that BMSC-derived exosomes regulate VC via the miR-335/SP1 axis, offering novel molecular targets for treating VC.