Altersbedingte Makuladegeneration: die Rolle von Licht bei der Entstehung degenerativer Veränderungen im menschlichen RPE und möglicher Zell-Schutz durch Minocyclin

Zusammenfassung

Hintergrund: Lichtinduzierter oxidativer Stress wird als mögliche Ursachen degenerativer Veränderungen im retinalen Pigmentepithel (RPE) bei der altersabhängigen Makuladegeneration (AMD) angesehen. Diese Studie untersucht den Einfluss von Licht auf die Induktion von intrazellulären freien Sauerstoffradikalen (ROS) und Apoptose im menschlichen RPE sowie mögliche anti-apoptotische, zytoprotektive Wirkungen des Tetrazyklin-Antibiotikums Minocyclin. Methoden: Kulturen primärer humaner RPE-Zellen wurden mit weißem Licht bestrahlt oder oxidativem Stress ausgesetzt (600 µM H ₂O₂) und zusätzlich mit Minocyclin behandelt. Es wurden Vitalität, Induktion von Apoptose und intrazellulären ROS sowie die Expression von BAX und Bcl-2 auf zellulärer Ebene bestimmt (RT-PCR/Western Blot/ELISA). Ergebnisse: Sowohl die Bestrahlung der RPE Zellen mit weißem Licht als auch die H ₂O₂-Exposition führten zu einer bestrahlungszeitabhängigen Abnahme der Vitalität und der Expression von Bcl-2 sowie zu einer Induktion von Apoptose, ROS und der Bildung von BAX. Diese Zellschädigung wurde unter bestimmten Bedingungen durch Behandlung der Zellen mit Minocyclin reduziert. Schlussfolgerung: Diese Studie zeigt, dass Minocyclin humane RPE-Zellen wirkungsvoll vor oxidativem Stress schützt. Aufgrund seiner photosensibilisierenden Wirkung muss die mögliche Anwendung von Minocyclin bei AMD aber weiter evaluiert werden.

Abstract

Background: Light-induced oxidative stress is an suggested reason for retinal pigment epithelium (RPE) degeneration in age-related macular degeneration (AMD). This study investigates the influence of light on intracellular reactive oxygen species (ROS) and apoptosis in the human RPE and potential cytoprotective effects of the tetracycline antibiotic minocycline. Methods: Primary human RPE cells were either pre- or post-incubated with minocycline and then exposed to white light or oxidative stress (600 µM, H ₂O₂). Then viability, induction of intracellular reactive oxygen species (ROS), apoptosis and cell death was determined. Expression of apoptotic BAX and anti-apoptotic Bcl-2 protein and their mRNA were determined by RT-PCR and Western blot analysis. Results: Both light exposure and oxidative stress decreased RPE cell viability and Bcl-2 expression and increased intracellular ROS, apoptotic cell death, and BAX expression. Minocycline reduced these effects under certain conditions. Conclusions: This study demonstrates that minocycline effectively protects human RPE cells against oxidative damage. However, in the light of minocycline’s photosensitising properties its potential role in AMD treatment needs further evaluation.

Schlüsselwörter

oxidativer Stress - Minocyclin - AMD - Apoptose - Zytoprotektion

Key words

oxidative stress - Pharmacology - AMD - apoptosis - minocycline

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