眼内注射吲哚菁绿对视网膜细胞有毒性。

Intravitreal indocyanine green is toxic to the retinal cells.

机构信息

Department of Pharmacy, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200032, China; Department of Ophthalmology, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China.

Department of Ophthalmology, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China; The International Eye Research Institute of the Chinese University of Hong Kong (Shenzhen), Shenzhen, 518000, China; C-MER (Shenzhen) Dennis Lam Eye Hospital, Shenzhen, 518000, China; C-MER Dennis Lam & Partners Eye Center, C-MER International Eye Care Group, Hong Kong, 999077, China.

出版信息

Biochem Biophys Res Commun. 2024 Dec 3;736:150872. doi: 10.1016/j.bbrc.2024.150872. Epub 2024 Oct 22.

DOI:10.1016/j.bbrc.2024.150872

PMID:39471681

Abstract

BACKGROUND

Indocyanine green (ICG) is widely used to stain the epiretinal membranes and internal limiting membranes during the pars plana vitrectomy (PPV). This study aims to evaluate the effect of ICG on rat retinas and various retinal cell lines, including ARPE-19 cells, rMC-1 cells, BV2 cells, HRMECs and R28 cells.

METHODS

ICG solutions were prepared and diluted with glucose solution (GS) according to the standard clinical protocols. The retinal cell lines, including ARPE-19 cells, rMC-1 cells, BV2 cells, HRMECs and R28 cells, were treated with the following solutions: normal glucose (NG, 5 mM), GS-1 (92.5 mM glucose), GS-2 (185.02 mM glucose), ICG-1 (92.5 mM glucose + 0.43 mM ICG), or ICG-2 (185.02 mM glucose + 0.86 mM ICG) for durations of 15 or 30 min. In vivo, the right eyes of the rats were intravitreally injected with ICG-1 or ICG-2 (2 μL), while the left eyes were intravitreally injected with GS-1 or GS-2, served as the osmotic controls, for 30 min or 60 min. The rats intravitreally injected with an equivalent volume of NG or 1x phosphate-buffered saline (1x PBS) were served as the normal control or vehicle control. The cell viability was measured with the Cell Counting Kit-8 (CCK-8), while the cell death in retinal cryosections was detected with the TUNEL assay.

RESULTS

The viabilities of the different retinal cell lines involved in this study were significantly reduced by both ICG-1 and ICG-2 treatments at both time points, with ICG-2 resulting in lower cell viability compared to the NG group and the osmotic control group. Additionally, GS-2 treatment also exhibited a decrease in retinal cell viabilities in vitro. To further confirm these results, intravitreal injection of ICG or GS induced more apoptotic cell death in rat retinas as evidenced by the TUNEL assay.

CONCLUSIONS

The exposure of ICG or its solvent leads to an augmented retinal cell death, which is directly proportional to the concentration and duration of exposure, both in vivo and in vitro. Caution should be exercised during vitrectomy procedures involving ICG administration during clinical practice. It is recommended to advocate for lower concentrations of ICG with reduced exposure time during ocular surgeries.

摘要

背景

吲哚菁绿（ICG）在经睫状体平坦部玻璃体切除术（PPV）中广泛用于染色视网膜内界膜和视网膜外膜。本研究旨在评估 ICG 对大鼠视网膜和各种视网膜细胞系的影响，包括 ARPE-19 细胞、rMC-1 细胞、BV2 细胞、HRMECs 和 R28 细胞。

方法

根据标准临床方案，用葡萄糖溶液（GS）制备和稀释 ICG 溶液。将 ARPE-19 细胞、rMC-1 细胞、BV2 细胞、HRMECs 和 R28 细胞等视网膜细胞系分别用以下溶液处理：正常葡萄糖（NG，5mM）、GS-1（92.5mM 葡萄糖）、GS-2（185.02mM 葡萄糖）、ICG-1（92.5mM 葡萄糖+0.43mM ICG）或 ICG-2（185.02mM 葡萄糖+0.86mM ICG），处理时间为 15 或 30min。在体内，将 ICG-1 或 ICG-2（2μL）玻璃体腔内注射到大鼠右眼，左眼玻璃体腔内注射 GS-1 或 GS-2，作为渗透对照，持续 30 或 60min。将等效体积的 NG 或 1x 磷酸盐缓冲盐水（1x PBS）玻璃体腔内注射到大鼠右眼作为正常对照或载体对照。用细胞计数试剂盒-8（CCK-8）测量细胞活力，用 TUNEL 测定法检测视网膜冷冻切片中的细胞死亡。

结果

在这两个时间点，ICG-1 和 ICG-2 处理均显著降低了本研究中涉及的不同视网膜细胞系的活力，与 NG 组和渗透对照组相比，ICG-2 导致更低的细胞活力。此外，GS-2 处理也导致体外视网膜细胞活力下降。为了进一步证实这些结果，用 TUNEL 测定法证实，玻璃体腔内注射 ICG 或 GS 诱导大鼠视网膜中更多的细胞凋亡。