Memantine, Simvastatin, and Epicatechin Inhibit 7-Ketocholesterol-induced Apoptosis in Retinal Pigment Epithelial Cells But Not Neurosensory Retinal Cells In Vitro

Overview

Journal J Ophthalmic Vis Res

Specialty Ophthalmology

Date 2020 Nov 2

PMID 33133437

Citations 2

Authors

Aneesh Neekhra

Julia Tran

Parsa R Esfahani

Kevin Schneider

Khoa Pham

Ashish Sharma

Marilyn Chwa

Saurabh Luthra

Ana L Gramajo

Saffar Mansoor

Baruch D Kuppermann

M Cristina Kenney

Affiliations

Soon will be listed here.

Abstract

Purpose: 7-ketocholesterol (7kCh), a natural byproduct of oxidation in lipoprotein deposits is implicated in the pathogenesis of diabetic retinopathy and age-related macular degeneration (AMD). This study was performed to investigate whether several clinical drugs can inhibit 7kCh-induced caspase activation and mitigate its apoptotic effects on retinal cells in vitro.

Methods: Two populations of retinal cells, human retinal pigment epithelial cells (ARPE-19) and rat neuroretinal cells (R28) were exposed to 7kCh in the presence of the following inhibitors: Z-VAD-FMK (pan-caspase inhibitor), simvastatin, memantine, epicatechin, and Z-IETD-FMK (caspase-8 inhibitor) or Z-ATAD-FMK (caspase-12 inhibitor). Caspase-3/7, -8, and -12 activity levels were measured by fluorochrome caspase assays to quantify cell death. IncuCyte live-cell microscopic images were obtained to quantify cell counts.

Results: Exposure to 7kCh for 24 hours significantly increased caspase activities for both ARPE-19 and R28 cells ( 0.05). In ARPE cells, pretreatment with various drugs had significantly lower caspase-3/7, -8, and -12 activities, reported in % change in mean signal intensity (msi): Z-VAD-FMK (48% decrease, 0.01), memantine (decreased 47.8% at 1 µM, = 0.0039 and 81.9% at 1 mM, 0.001), simvastatin (decreased 85.3% at 0.01 µM, 0.001 and 84.8% at 0.05 µM, 0.001) or epicatechin (83.6% decrease, 0.05), Z-IETD-FMK (68.1% decrease, 0.01), and Z-ATAD-FMK (47.7% decrease, = 0.0017). In contrast, R28 cells exposed to 7kCh continued to have elevated caspase-3/7, -8, and -12 activities (between 25.7% decrease and 17.5% increase in msi, 0.05) regardless of the pretreatment.

Conclusion: Several current drugs protect ARPE-19 cells but not R28 cells from 7kCh-induced apoptosis, suggesting that a multiple-drug approach is needed to protect both cells types in various retinal diseases.

Citing Articles

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