Sheu Shwu-Jiuan, Chen Jiunn-Liang, Bee Youn-Shen, Chen Yi-An, Lin Shi-Han, Shu Chih-Wen
Department of Ophthalmology, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan.
School of Medicine, National Yang-Ming University, Taipei, Taiwan.
PLoS One. 2017 Mar 30;12(3):e0174736. doi: 10.1371/journal.pone.0174736. eCollection 2017.
Indocyanine green (ICG) and brilliant blue G (BBG) are commonly used vital dyes to remove internal limiting membrane (ILM) in vitreoretinal surgery. The vital dyes have shown cytotoxic effects in ocular cells. Autophagy is a stress responsive pathway for either protecting cells or promoting cell death. However, the role of autophagy in ocular cells in response to the vital dyes remains unknown. In this study, we found that ICG and BBG reduced cell viability in both human retinal pigment epithelial ARPE-19 and mouse photoreceptor 661W cells. ICG and BBG induced lipidated GFP-LC3-II and LC3-II in ARPE-19 and 661W cells. Combination treatment with the autophagy inhibitor chloroquine indicated that ICG and BBG reduced autophagic flux in ARPE-19 cells, whereas the vital dyes induced autophagic flux in 661W cells. Moreover, genetic and pharmacological ablation of autophagy enhanced vital dyes-induced cytotoxicity in ocular cells. Dietary supplements, including resveratrol, lutein, and CoQ10, induced autophagy and diminished the cytotoxic effects of ICG and BBG in ocular cells. These results suggest that autophagy may protect ARPE-19 and 661W cells from vital dyes-induced damage.
吲哚菁绿（ICG）和亮蓝G（BBG）是玻璃体视网膜手术中常用的用于去除内界膜（ILM）的活性染料。这些活性染料已在眼细胞中显示出细胞毒性作用。自噬是一种应激反应途径，可保护细胞或促进细胞死亡。然而，自噬在眼细胞中对活性染料的反应作用仍不清楚。在本研究中，我们发现ICG和BBG降低了人视网膜色素上皮ARPE - 19细胞和小鼠光感受器661W细胞的细胞活力。ICG和BBG在ARPE - 19和661W细胞中诱导了脂化的GFP - LC3 - II和LC3 - II。自噬抑制剂氯喹的联合处理表明，ICG和BBG降低了ARPE - 19细胞中的自噬通量，而活性染料在661W细胞中诱导了自噬通量。此外，自噬的基因和药理学消融增强了活性染料在眼细胞中诱导的细胞毒性。包括白藜芦醇、叶黄素和辅酶Q10在内的膳食补充剂诱导了自噬，并减弱了ICG和BBG在眼细胞中的细胞毒性作用。这些结果表明，自噬可能保护ARPE - 19和661W细胞免受活性染料诱导的损伤。
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