Yiğit Musa, Güneş Alime, Uğuz Cihangir, Yalçın Tök Özlem, Tök Levent, Öz Ahmi, Nazıroğlu Mustafa
Department of Ophthalmology, Süleyman Demirel University Research and Education Hospital, Çünür-Isparta 32200, Turkey.
Department of Biophysics, Faculty of Medicine, Süleyman Demirel University Research and Education Hospital, Çünür-Isparta 32200, Turkey.
Int J Ophthalmol. 2019 Jun 18;12(6):930-935. doi: 10.18240/ijo.2019.06.08. eCollection 2019.
To observe the protective effect of astaxanthin (AST) against hydroquinone (HQ) mediated cell death in the apoptotic cascade and evaluate intracellular Ca release, caspase-3, and -9 activation, reactive oxygen species (ROS) production in ARPE-19 cells.
We cultured ARPE-19 cells in special mediums and performed MTT tests to determine protective effect of AST, before exposing the cells to HQ in an incubator. We analyzed intracellular Ca release experiments, mitochondrial membrane depolarization, glutathione (GSH), glutathione peroxidase (GSH-Px) and ROS experiments, and apoptosis assay.
ROS production ranges depend on the amount of cell death. We computed the correlation between ROS ranges and cell death by 20,70-dichlorofluorescein fluorescence, and Ca levels by Fura-2-AM. HQ-induced cell death found out to rise ranges of caspase-3 and -9, and mitochondrial depolarization. These three steps were delayed by AST management.
ARPE-19 cells are avoided from HQ-induced ROS production and caspase-3 and -9 activation by AST. AST may limit the range of caspase synthesis, Ca release and excess production of ROS with antiapoptotic effect. This study proposes a new therapeutic approach for the treatment of age-related macular degeneration.
观察虾青素（AST）对苯二酚（HQ）介导的细胞凋亡级联反应中细胞死亡的保护作用，并评估ARPE-19细胞内钙释放、半胱天冬酶-3和-9的激活以及活性氧（ROS）的产生。
我们在特殊培养基中培养ARPE-19细胞，并进行MTT试验以确定AST的保护作用，然后将细胞置于培养箱中暴露于HQ。我们分析了细胞内钙释放实验、线粒体膜去极化、谷胱甘肽（GSH）、谷胱甘肽过氧化物酶（GSH-Px）和ROS实验以及凋亡检测。
ROS产生范围取决于细胞死亡量。我们通过2′,7′-二氯荧光素荧光计算ROS范围与细胞死亡之间的相关性，通过Fura-2-AM计算钙水平。发现HQ诱导的细胞死亡会增加半胱天冬酶-3和-9的范围以及线粒体去极化。这三个步骤通过AST处理而延迟。
AST可使ARPE-19细胞避免HQ诱导的ROS产生以及半胱天冬酶-3和-9的激活。AST可能通过抗凋亡作用限制半胱天冬酶合成、钙释放和ROS过量产生的范围。本研究提出了一种治疗年龄相关性黄斑变性的新治疗方法。
