MicroRNA-100 Mediates Hydrogen Peroxide-Induced Apoptosis of Human Retinal Pigment Epithelium ARPE-19 Cells

Abstract

This study investigated the regulatory role of microRNA 100 (miR-100) in hydrogen peroxide (H2O2)-induced apoptosis of human retinal pigment epithelial ARPE-19 cells. H2O2 induced oxidative cell death of cultured ARPE-19 cells was measured by cytotoxicity assay. qRT-PCR was used to quantify cytosolic and extracellular contents of miR-100. Kinase and miR-100 inhibition treatments were applied to determine the regulatory signaling pathways involved in cell death regulation. H2O2 dose-dependently reduced viability of ARPE-19 cells and simultaneously upregulated miR-100 levels in both cytosolic and extracellular compartments. Western blotting detection indicated that H2O2 elicited hyperphosphorylation of PI3K/Akt, ERK1/2, JNK, p38 MAPK, and p65 NF-κB. Further kinase inhibition experiments demonstrated that PI3K, p38 MAPK, and NF-κB activities were involved in oxidative-stress-induced miR-100 upregulation in ARPE-19 cells, while blockade of PI3K, JNK, and NF-κB signaling significantly attenuated the oxidative cell death. Intriguingly, MiR-100 antagomir treatment exerted a cytoprotective effect against the H2O2-induced oxidative cell death through attenuating the oxidation-induced AMPK hyperphosphorylation, restoring cellular mTOR and p62/SQSTM1 levels and upregulating heme oxygenase-1 expression. These findings support that miR-100 at least in part mediates H2O2-induced cell death of ARPE-19 cells and can be regarded as a preventive and therapeutic target for retinal degenerative disease.

MiR-6837-3p protected retinal epithelial cells from oxidative stress by targeting E2F6 Running Title：MiR-6837-3p Protected retinal cells

“…In AMD disease, RPE cells may experience oxidative stress-induced senescence. Whereas oxidative stress products such as H 2 O 2 attack DNA, proteins, lipids, and other cellular components indiscriminately and plays an important role in controlling cellular functions such as cell differentiation, proliferation, migration, apoptosis, and death (4,5) MicroRNA (miRNA) can inhibit endogenous gene expression by translationally cleaving gene transcripts (9). It can regulate the biological functions of cells by targeting several downstream genes, thus playing an important regulatory role in various diseases, including AMD.…”

“…Aberrant expression of let-7b, let-7a, miR-125b, miR-24, miR-320, miR-23b, let-7e, and let-7d miRNAs in ROS-accumulating ARPE-19 cells was determined by microarray hybridization analysis, which caused apoptosis and resulted in an increase in the expression of the target genes HMOX1 and GADD153 (13). It has also been shown that treatment with miR-100 antagomir prevented excessive oxidation-induced cell death in ARPE-19 cells and exerts a protective effect on oxidative cell death (5).…”

“…Oxidative stress leads to excessive production of reactive oxygen species (ROS), including superoxide, hydroxyl radicals, and hydrogen peroxide (H 2 O 2 ), which can cause severe oxidative damage to retinal pigment epithelial (RPE) cells, retinal ganglion cells (RGCs), and others (1), and this is the main pathomechanism of age-related macular degeneration (AMD) and other retinal degenerative diseases (2,3) ROS such as H 2 O2 attack DNA, proteins, lipids and other cellular components indiscriminately and plays an important role in controlling cellular functions such as cell differentiation, proliferation, migration, apoptosis and death (4,5). It has been shown that oxidative stress induces the expression of pentraxin 3 in RPE cells, which is involved in RPE cells dysfunction(6).…”

Immunomodulatory and Antioxidant Drugs in Glaucoma Treatment

“…The activation of the apoptosis signal-regulating kinase 1 (ASK-1)/p38 mitogen-activated protein kinase (MAPK)/JNK/extracellular-signal-regulated kinase (ERK) axis by ROS can lead to caspase-3 activation and cellular membrane disassembly, promoting cell death [ 99 , 100 ]. Chronic exposure to ROS can activate the phosphoinositide 3-kinase (PI3K)/Akt axis while attenuating the mammalian target of rapamycin (mTOR) pathway, further stimulating the NF-kB and enhancing inflammatory events [ 101 ]. Additionally, excessive ROS disrupts glutamate metabolism, leading to the neurotoxic extracellular accumulation of glutamate, as dysfunctional glial cells fail to properly buffer the excess glutamate [ 16 , 102 , 103 ].…”

Oxidative Stress: A Suitable Therapeutic Target for Optic Nerve Diseases?

“…This axis triggers the mitochondrial apoptotic pathway, characterized by the release of the Bcl-2 associated X-protein (Bax) and cytochrome c into the cytosol, followed by apoptosome formation (cytochrome c/apoptotic protease-activating factor 1(Apaf-1)/caspase-9), ultimately leading to caspase-3 activation [220,222,223]. Moreover, hydrogen peroxide has been described to activate the phosphoinositide 3-kinase (PI3K)/ Ak strain transforming (Akt) axis and reduce the intracellular concentration of the mammalian target of rapamycin (mTor), promoting NF-kB activation [224].…”