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.

Highlights

  • Recent studies have evidenced that the existence of miRNA in the extracellular vesicles released by retinal pigment epithelium (RPE) cells may contribute to retinal pathophysiolgical processes [35,36]

  • The results indicated that both cytosolic and released contents of microRNA 100 (miR-100) significantly increased in a dose-dependent manner by oxidative stress (Figure 1B,C)

  • Given that the AMPK-dependent upregulation of heme oxygenase-1 (HO-1) antioxidant expression protects ARPE-19 cells against oxidative-stress-induced apoptosis [42], we examined whether AMPK/mTOR axis and HO-1 expression are involved in the miR-100 antagomir exhibited cytoprotection against oxidative injury of RPE cells

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Summary

Introduction

Age-related macular degeneration (AMD) is the most common cause of blindness among patients aged 65 years and older in the Western world [1], and incidence continues to rise due to the increasing percentage of older adults in the common population. In. Asia, the prevalence of late AMD is estimated to be between 0.20 and 1.90%, based on data from China [2], Japan [3], Taiwan [4], and Singapore [5]. AMD results from retinal pigment epithelium (RPE) dysfunction or loss associated with photoreceptor fallout, Bruch’s membrane thickening, and choriocapillary hypoperfusion [6]. The RPE is a Pharmaceuticals 2021, 14, 314.

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