MiR-125b-5p ameliorates ox-LDL-induced vascular endothelial cell dysfunction by negatively regulating TNFSF4/TLR4/NF-κB signaling.

Publisher: BioMed Central Country of Publication: England NLM ID: 101088663 Publication Model: Electronic Cited Medium: Internet ISSN: 1472-6750 (Electronic) Linking ISSN: 14726750 NLM ISO Abbreviation: BMC Biotechnol Subsets: MEDLINE

Original Publication: [London] : BioMed Central, 2001-

Lipoproteins, LDL*/pharmacology ; Lipoproteins, LDL*/metabolism ; MicroRNAs*/genetics ; MicroRNAs*/metabolism ; Toll-Like Receptor 4*/metabolism ; Toll-Like Receptor 4*/genetics ; Human Umbilical Vein Endothelial Cells*/metabolism ; Signal Transduction*/drug effects ; NF-kappa B*/metabolism ; OX40 Ligand*/metabolism ; OX40 Ligand*/genetics; Humans ; Apoptosis/drug effects ; Oxidative Stress/drug effects ; Atherosclerosis/metabolism ; Atherosclerosis/genetics

Background: Oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell dysfunction plays a crucial role in the progression of atherosclerosis (AS). Although miR-125b-5p is known to be involved in cardiovascular and cerebrovascular disorders, its function in ox-LDL-induced endothelial injury is still not well understood.
Methods: An in vitro AS cell model was established by exposing human umbilical vein endothelial cells (HUVECs) to 100 µg/mL ox-LDL for 24 h. A series of functional assays, including CCK-8 assay, flow cytometry, MDA and SOD kits, capillary-like network formation assay and ELISA assay were performed in vitro. TNFSF4/TLR4/NF-κB pathway-related protein expressions were measured by Western blot. Molecular mechanisms were elucidated through quantitative real-time PCR, western blot analysis, and luciferase reporter assays.
Results: Our investigation revealed that exposure to ox-LDL led to a downregulation in miR-125b-5p, while upregulating the expression of tumor necrosis factor (ligand) superfamily, member 4 (TNFSF4), TLR4, p-p65 and p-IkBa in HUVECs in a dose-dependent manner. We confirmed TNFSF4 as a direct target of miR-125b-5p. Ox-LDL exposure led to decreased cell viability and angiogenic capacity, along with increased apoptosis, inflammation, and oxidative stress in HUVECs. These effects were reversed by overexpressing miR-125b-5p or knocking down TNFSF4. Overexpression of TNFSF4 significantly reversed the effects brought about by miR-125b-5p in HUVECs exposed to ox-LDL. Moreover, miR-125b-5p inactivated the TLR4/NF-κB signaling pathway by negatively regulating TNFSF4.
Conclusions: In summary, our findings demonstrate that miR-125b-5p possessed an anti-inflammatory and anti-apoptosis against ox-LDL-induced HUVEC injury by regulating the TNFSF4/TLR4/NF-κB signaling, indicating that miR-125b-5p may have an important therapeutic function for AS.
Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests.
(© 2025. The Author(s).)

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Keywords: Atherosclerosis; HUVECs; TLR4/NF-κB signaling; TNFSF4; miR-125b-5p

0 (Lipoproteins, LDL)
0 (oxidized low density lipoprotein)
0 (MicroRNAs)
0 (Toll-Like Receptor 4)
0 (MIRN125 microRNA, human)
0 (NF-kappa B)
0 (TLR4 protein, human)
0 (OX40 Ligand)
0 (TNFSF4 protein, human)

Date Created: 20250125 Date Completed: 20250125 Latest Revision: 20250130

20250130

PMC11760099

10.1186/s12896-025-00944-y

39856662