miR-486-5p Restrains Extracellular Matrix Production and Oxidative Damage in Human Trabecular Meshwork Cells by Targeting TGF-β/SMAD2 Pathway

Abstract

Background. Glaucoma is characterized by elevated intraocular pressure caused by aqueous outflow dysfunction. Trabecular meshwork plays a key role in controlling intraocular pressure by modulating aqueous outflow. This study investigated the protective effects of miR-486-5p in H2O2-stimulated human trabecular meshwork cells (TMCs). Methods. TMCs were disposed with 300 μM H2O2 to establish oxidative damage models in vitro. miR-486-5p mimics and its controls were transfected into TMCs, and cell apoptosis and extracellular matrix production (ECM) genes were measured by flow cytometry, western blotting, and immunofluorescence staining. Activities of superoxide dismutase (SOD) and malondialdehyde (MDA) were also assayed. Online tools and luciferase reporter assays were used to uncover the relationship between miR-486-5p and the TGF-β/SMAD2 pathway. Results. We found that H2O2-induced oxidative damage in TMCs and miR-486-5p was downregulated in H2O2-stimulated TMCs. Overexpression of miR-486-5p mitigated H2O2-induced oxidative damage by inhibiting apoptosis, reducing cleaved caspase-3/9 expression, reducing MDA levels, and increasing SOD levels as well as downregulating ECM genes. SMAD2 was demonstrated to be targeted by miR-486-5p, and miR-486-5p inhibited TGF-β/SMAD2 signaling in H2O2-stimulated TMCs. Additionally, SMAD2 was upregulated by H2O2, and SMAD2 upregulation abrogated the protective effects of miR-486-5p against H2O2-induced injury. Conclusion. miR-486-5p restrains H2O2-induced oxidative damage in TMCs by targeting the TGF-β/SMAD2 pathway.

S-RBD-modified and miR-486-5p-engineered exosomes derived from mesenchymal stem cells alleviate radiation-induced lung injury and long-term pulmonary fibrosis via suppression of ferroptosis

“…Several important signaling regulators, such as PTEN, FOXO1, and SIRT1, have been identi ed as targets of miR-486-5p [14]. Extensive studies have revealed that miR-486-5p is involved in the regulation of resistance to oxidative damage and the inhibition of collagen hyperproliferation, and brosis [44][45][46]. Downregulated expression of miR-486-5p has been observed in the serum and lung tissues of patients with silicosis and idiopathic pulmonary brosis [18].…”

Glaucoma: Novel antifibrotic therapeutics for the trabecular meshwork

“…MiR-486–5p is a plasma and extracellular vesicle-enriched miRNA ( Douvris et al, 2022 ). As shown by the results of small RNA sequencing, miR-486–5p was downregulated in POAG patients ( Hubens et al, 2021 ), and transfection of miR-486–5p mimics in TM cells can significantly inhibit H 2 O 2 -stimulated ECM remodeling by blocking the TGF-β2/Smad2 signaling pathway ( Xu et al, 2022 ). MiR-483 generally exists in two mature forms, miR-483–3p and miR-483–5p, in the body and is closely related to cell proliferation and tumor development ( Watson et al, 2013 ; Zhou et al, 2018 ).…”

“… miR-29 human TM cell ( Luna et al, 2009 ; Villarreal et al, 2011 ) Suppressing various EMC, like SPARC, collagen I, collagen IV, and laminin expression and negatively regulate genes involved in ECM deposition and remodeling. miR-486–5p human TM cell ( Xu et al, 2022 ) Inhibiting H 2 O 2 -stimulated ECM remodeling by blocking TGF-β2/Smad2 signaling pathway. miR-483–3p human TM cell ( Shen et al, 2015 ) Protecting TM cell from 300 μM H 2 O 2 induced-fibrosis by inhibiting TGF-β2/Smad4 signaling pathway.…”

The Application of MicroRNAs in Glaucoma Research: A Bibliometric and Visualized Analysis

“…Therefore, miR-200c is a valuable candidate for future therapeutic approaches in glaucoma research by regulating TM cell contraction. In addition, miR-486-5p can reduce the production of ECM and oxidative damage in human TM cells by targeting the TGF-β/SMAD2 pathway [101]. Overall, future miRNA-based therapies that focus on controlling the production of ECM, regulating TM cell contraction, or targeting the TGF-β may provide novel therapeutic approaches for glaucoma.…”