Wu Juan, Gao Zhen-Ya, Cui Dong-Mei, Li Hong-Hui, Zeng Jun-Wen
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, Guangdong Province, China.
Xuchang University, School of Medicine, Xuchang 461000, Henan Province, China.
Int J Ophthalmol. 2020 Sep 18;13(9):1345-1350. doi: 10.18240/ijo.2020.09.01. eCollection 2020.
To explore the apoptosis of ARPE-19 cells after the treatment with different doses of all-trans-retinoic acid (ATRA).
ARPE-19 cells were used in the experiment. Flow cytometry assay was employed to evaluate the level of reactive oxygen species (ROS) and apoptosis. The effects of ATRA (concentrations from 2.5 to 20 µmol/L) on the expression of endoplasmic reticulum stress (ERS) markers were evaluated by Western blot and real-time quantitative polymerase chain reaction (qRT-PCR) assays. The contribution of ROS and ERS-induced apoptosis was determined by using N-acetyl-L-cysteine (NAC) and Salubrinal, an antagonist of NAC and ERS, respectively.
Flow cytometry showed that ATRA significantly increased ARPE-19 cell apoptosis and ROS levels in each group (=86.39, <0.001; =116.839, <0.001). Western blot and qRT-PCR revealed that levels of CHOP and BIP were elevated in a concentration-dependent pattern after the cells were incubated with ATRA (2.5-20 µmol/L). The upregulation of VEGF-A and CHOP induced by ATRA could be inhibited by NAC (antioxidant) and Salubrinal (ERS inhibitor) .
ATRA induces the apoptosis of ARPE-19 cells activated ROS and ERS signaling pathways.
探讨不同剂量全反式维甲酸（ATRA）处理后ARPE - 19细胞的凋亡情况。
实验采用ARPE - 19细胞。运用流式细胞术检测活性氧（ROS）水平及细胞凋亡情况。通过蛋白质免疫印迹法（Western blot）和实时定量聚合酶链反应（qRT - PCR）检测ATRA（浓度为2.5至20 μmol/L）对内质网应激（ERS）标志物表达的影响。分别使用N - 乙酰 - L - 半胱氨酸（NAC）和ERS拮抗剂Salubrinal确定ROS和ERS诱导细胞凋亡的作用。
流式细胞术显示，ATRA显著增加了各组ARPE - 19细胞凋亡率和ROS水平（F = 86.39，P < 0.001；F = 116.839，P < 0.001）。蛋白质免疫印迹法和qRT - PCR显示，用ATRA（2.5 - 20 μmol/L）孵育细胞后，CHOP和BIP水平呈浓度依赖性升高。ATRA诱导的VEGF - A和CHOP上调可被NAC（抗氧化剂）和Salubrinal（ERS抑制剂）抑制。
ATRA通过激活ROS和ERS信号通路诱导ARPE - 19细胞凋亡。
