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MiR-375 Inhibitor Alleviates Inflammation and Oxidative Stress by Upregulating the GPR39 Expression in
Abstract:
Atherosclerosis may be caused or developed by an immune response and antioxidation imbalance. MicroRNA-375 (miR-375) or G-protein-coupled receptor 39 (GPR39) is involved in vascular endothelial cell injury, but their role in atherosclerosis is unknown. This experiment aimed to determine the action of the miR-375/GPR39 axis in atherosclerosis.Human aortic endothelial cells (HAECs) were treated with 10 ng/mL of oxidised low-density lipoprotein (ox-LDL) for 24 hours to induce HAEC injury, which was treated by the miR-375 inhibitor, GPR39 inhibitor, or agonist. High-fat diet (HFD) -induced ApoE-/- mice were made as an atherosclerosis model for miR-375 inhibitor treatment. Cell Counting Kit-8 was applied to detect HAEC viability. HAEC apoptosis and ROS levels were measured using flow cytometry. Vascular histopathology and the GPR39 expression were detected using hematoxylin-eosin and immunohistochemistry. The expressions of interleukin (IL) -6, IL-1β, and tumour necrosis factor-α (TNF-α) were assessed using an enzyme-linked immunosorbent assay. The miR-375, GPR39, NOX-4, and p-IκBα/IκBα levels were measured using quantitative reverse transcription polymerase chain reaction or western blot.MiR-375 and GPR39 levels increased and decreased in ox-LDL-treated HAECs, respectively. The miR-375 inhibitor or GPR39 agonist promoted cell viability and inhibited apoptosis in ox-LDL-induced HAEC injury. The miR-375 inhibitor also significantly downregulated the IL-6, IL-1β, TNF-α, p-IκBα/IκBα, ROS, and NOX-4 expressions to alleviate oxidative stress and inflammation, which were reversed by the GPR39 inhibitor. An in vivo experiment proved that the miR-375 inhibitor upregulated the GPR39 expression and improved inflammation, oxidative stress, and endothelial cell damage associated with atherosclerosis.The miR-375 inhibitor improved inflammation, oxidative stress, and cell damage in ox-LDL-induced HAECs and HFD-induced ApoE-/- mice by promoting the GPR39 expression, which provided a new theoretical basis for the clinical treatment of atherosclerosis.
Insights
The miR-375 inhibitor improved atherosclerosis by upregulating G-protein-coupled receptor 39 (GPR39), reducing inflammation and oxidative stress in endothelial cells. This finding offers a new therapeutic strategy for atherosclerosis treatment.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Immunology
Background:
- Atherosclerosis pathogenesis involves immune responses and oxidative stress.
- MicroRNA-375 (miR-375) and G-protein-coupled receptor 39 (GPR39) are implicated in endothelial cell injury.
- The specific roles of the miR-375/GPR39 axis in atherosclerosis remain unclear.
Purpose of the Study:
- To investigate the role of the miR-375/GPR39 axis in atherosclerosis.
- To determine the therapeutic potential of targeting this axis in atherosclerosis models.
Main Methods:
- Human aortic endothelial cells (HAECs) were exposed to oxidized low-density lipoprotein (ox-LDL) to model endothelial injury.
- Treatments included miR-375 inhibitor, GPR39 inhibitor, or agonist.
- Atherosclerosis was induced in ApoE-/- mice using a high-fat diet (HFD).
- Cell viability, apoptosis, reactive oxygen species (ROS), inflammatory cytokines (IL-6, IL-1β, TNF-α), and molecular markers (GPR39, NOX-4, p-IκBα/IκBα) were assessed.
Main Results:
- Oxidized low-density lipoprotein (ox-LDL) increased miR-375 and decreased GPR39 levels in HAECs.
- miR-375 inhibition or GPR39 agonism enhanced HAEC viability and reduced apoptosis.
- The miR-375 inhibitor downregulated inflammatory markers, ROS, and oxidative stress markers, an effect reversed by the GPR39 inhibitor.
- In vivo studies confirmed that miR-375 inhibition upregulated GPR39, mitigating atherosclerosis-related inflammation, oxidative stress, and endothelial damage.
Conclusions:
- The miR-375 inhibitor ameliorates ox-LDL-induced endothelial injury and HFD-induced atherosclerosis in ApoE-/- mice.
- This protective effect is mediated by promoting GPR39 expression, thereby reducing inflammation and oxidative stress.
- The miR-375/GPR39 axis represents a potential therapeutic target for atherosclerosis.
