Abstract
Purpose: :
We have previously shown that oxidative stress (OS) can quantitatively regulate AP-1 gene expression changes, vitamin C by modulating AP-1 gene family played an important antioxidant role under OS in the human retinal pigment epithelium (RPE). The purpose of this study was to determine whether carnosic acid, genistein, quercetin, melatonin, and taurine pretreatment can quantitatively modulate these molecular responses respectively under OS.
Methods: :
Confluent ARPE-19 cells were cultured for three days in defined media in the presence of carnosic acid (0.5µg/ml - 25µg/ml), genistein ( 0.5µM-10µM),quercetin ( 2.5µM-10µM), taurine ( 1.0mM-20mM ) or melatonin (1.25µM-10µM) respectively, and then treated with 500µM H2O2. RNA was isolated using a no-rinse method at 0-,1-,4-, and 8-hours after OS and compared to untreated controls at each time point. FosB, cFos, and ATF3 gene expression was determined by real-time RT-PCR.
Results: :
Pretreatment with genistein 0.5µM cFos gene expression decreased 60%; with quercetin 2.5µM cFos gene expression reduced 46.4%; with melatonin 1.25µM cFos expression reduced 76.9% compared to the controls after 500µM H2O2 in one hour oxidative stress. Pretreatment with carnosic acid 5.0µM and 2.5µM cFos gene expression reduced 75.5% and 79.4%, FosB gene reduced 89.2% and 91.4%, ATF3 gene expression reduced 94.2% and 86.6% respectively in one hour OS. Pretreatment with taurine 1.0mM, 5.0mM and 20mM cFos expression reduced 51.7%,40.1% and 73.7%; FosB expression reduced 44.5%,52% and 58.8% in the one hour OS. Pretreatment with melatonin 1.25µM cFos expression reduced 76.9% in the one hour OS.
Conclusions: :
Carnosic acid, genistein, quercetin , taurine, and melatonin may modulate AP-1 gene family to play the antioxidative effects to protect RPE cells under OS in our study.
Keywords: retinal pigment epithelium • antioxidants • gene/expression