Tokyo Metropolitan Geriatric Hospital, Tokyo, Japan.
Retina. 2012 Mar;32(3):613-6. doi: 10.1097/IAE.0b013e31822091df.
To investigate the effects of Brilliant Blue G (BBG) on rat retinal ganglion cells (RGCs), both in vitro and in vivo.
In vitro, rat RGCs were purified by a 2-step immunopanning procedure, briefly exposed to BBG (250 mg/L), and irradiated with an endoilluminator for 15 minutes or incubated in the presence of BBG (concentration, 2-250 mg/L) without irradiation. The number of viable RGCs was counted after 3 days in culture. In vivo, after rats received an intravitreal injection of 3 μL of BBG (0.25 and 2.5 mg/L), the number of viable RGCs was examined by a 1,1'-dioctadecyl-1-3,3,3',3'-tetramethylindocarbocyanine perchlorate-retrolabeling technique.
In vitro, a brief exposure to BBG and irradiation did not affect RGC survival. However, BBG reduced the number of viable RGCs in a dose-dependent manner when the cells were exposed for 3 days. In vivo, when rats received an intravitreous injection of 0.25 mg/L BBG, the number of viable RGCs was not affected. The number of viable RGCs showed a tendency to decrease in the 2.5-mg/L group 14 days after the injection, but the difference was not significant compared with the number in the saline-treated control group.
Brilliant Blue G exerts no detectable detrimental effect on RGCs after short-time exposure, and no significant toxic effect even after a longer-time exposure in the current experimental setting.
研究亮蓝 G(BBG)对体外和体内大鼠视网膜神经节细胞(RGC)的影响。
体外,采用两步免疫淘选法纯化大鼠 RGC,短暂暴露于 BBG(250mg/L),并用内照器照射 15 分钟,或在无照射的情况下孵育于 BBG(浓度 2-250mg/L)。培养 3 天后计数存活的 RGC 数。体内,大鼠接受 3μL 玻璃体注射 BBG(0.25 和 2.5mg/L)后,用 1,1'-二辛基-3,3,3',3'-四甲基吲哚羰花青高氯酸盐逆行标记技术检测存活的 RGC 数。
体外,BBG 短暂暴露和照射均不影响 RGC 存活。然而,当细胞暴露 3 天时,BBG 以剂量依赖性方式减少存活的 RGC 数。体内,当大鼠接受玻璃体注射 0.25mg/L BBG 时,存活的 RGC 数不受影响。注射后 14 天,2.5mg/L 组存活的 RGC 数呈下降趋势,但与生理盐水处理对照组相比,差异无统计学意义。
在当前实验条件下,BBG 短时间暴露对 RGC 无明显有害作用,长时间暴露也无明显毒性作用。