Kanamori Akiyasu, Naka Maiko, Fukuda Masahide, Nakamura Makoto, Negi Akira
Department of Surgery, Division of Ophthalmology, Kobe University Graduate School of Medicine, Kobe, Japan.
Exp Eye Res. 2009 Mar;88(3):535-41. doi: 10.1016/j.exer.2008.11.012. Epub 2008 Dec 3.
We investigated whether latanoprost has a direct anti-apoptotic effect in retinal ganglion cell (RGC) line and RGCs in the rat. RGC-5 cells were induced to undergo apoptosis by serum deprivation and exogenous glutamate. The level of cell death with or without latanoprost acid was monitored by an XTT assay and by immunocytochemistry with activated caspase-3. Changes in the level of intracellular calcium ([Ca(2+)]i) were measured with fluo-4 fluorescence. The XTT assay revealed that latanoprost acid increased RGC-5 cell viability. Latanoprost acid significantly reduced caspase-3 positive cells and suppressed [Ca(2+)]i evoked by glutamate. U0126, a mitogen-activated protein/extracellular signal-regulated kinase 1 and 2 inhibitor, partially blocked the rescue effect of latnanoprost acid (p=0.013). In vivo, rat RGCs were degenerated by optic nerve crush. After topical instillation of latanoprost for 7days, RGCs labeled with fluorogold were significantly. Retinal flatmounts were subjected to terminal dUTP nick end labeling (TUNEL) staining to detect apoptotic cells. TUNEL-positive cells were significantly decreased in eyes with topically instilled latanoprost (p=0.015). These data suggest that latanoprost has an neuroprotective ability in RGCs.
我们研究了拉坦前列素是否对大鼠视网膜神经节细胞（RGC）系和RGC具有直接的抗凋亡作用。通过血清剥夺和外源性谷氨酸诱导RGC-5细胞发生凋亡。采用XTT法和活化的半胱天冬酶-3免疫细胞化学法监测有无拉坦前列素酸时的细胞死亡水平。用fluo-4荧光法测量细胞内钙（[Ca(2+)]i）水平的变化。XTT法显示拉坦前列素酸可提高RGC-5细胞活力。拉坦前列素酸显著减少半胱天冬酶-3阳性细胞，并抑制谷氨酸诱发的[Ca(2+)]i升高。U0126，一种丝裂原活化蛋白/细胞外信号调节激酶1和2抑制剂，部分阻断了拉坦前列素酸的挽救作用（p=0.013）。在体内，大鼠RGC因视神经挤压而退化。局部滴注拉坦前列素7天后，用荧光金标记的RGC显著减少。对视网膜平铺片进行末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色以检测凋亡细胞。局部滴注拉坦前列素的眼中TUNEL阳性细胞显著减少（p=0.015）。这些数据表明拉坦前列素对RGC具有神经保护能力。
