Journal Article

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

Ren, Maojia, He, Wenshuai, Han, Yunfei, Wang, Pengfei, Zhao, Limin

2025

Overview

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.

• Publisher: BioMed Central,Springer Nature B.V,BMC
• Subject: Angiogenesis / Apoptosis / Apoptosis - drug effects / Applied Microbiology / Arteriosclerosis / Assaying / Atherosclerosis / Atherosclerosis - metabolism / Biochemical Engineering / Biomedical Engineering/Biotechnology / Biotechnology / Capillary flow / Cardiovascular disease / Cell viability / Cells / Cerebrovascular diseases / Chemistry / Chemistry and Materials Science / Cholecystokinin / Cytokines / dose response / Endothelial cells / Enzyme-linked immunosorbent assay / Exposure / Flow cytometry / Genes / Genetic Engineering / Human Umbilical Vein Endothelial Cells - drug effects / Human Umbilical Vein Endothelial Cells - metabolism / Humans / HUVECs / In vitro methods and tests / inflammation / ligands / Lipoproteins, LDL - metabolism / Lipoproteins, LDL - pharmacology / Low density lipoprotein / luciferase / Membranes / MicroRNAs / MicroRNAs - genetics / MicroRNAs - metabolism / miR-125b-5p / Molecular modelling / Network formation / NF-kappa B - metabolism / NF-κB protein / OX40 Ligand - genetics / OX40 Ligand - metabolism / oxidation / Oxidative stress / Plant Breeding/Biotechnology / Proteins / quantitative polymerase chain reaction / Reagents / Real time / Signal transduction / Signal Transduction - drug effects / superfamily / therapeutics / TLR4 protein / TLR4/NF-κB signaling / TNFSF4 / Toll-Like Receptor 4 - genetics / Toll-Like Receptor 4 - metabolism / Toll-like receptors / Tumor necrosis factor-TNF / tumor necrosis factors / Umbilical vein / Western blotting