Tafluprost protects rat retinal ganglion cells from apoptosis in vitro and in vivo
Tafluprost protects rat retinal ganglion cells from apoptosis in vitro and in vivo
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DOI：
10.1007/s00417-009-1122-6
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发表时间：
2009-10-01
影响因子：
2.7
通讯作者：
Negi, Akira
中科院分区：
文献类型：
--
作者：
Kanamori, Akiyasu;Naka, Maiko;Negi, Akira
为了研究他氟前列素（一种与奥沙利铂相关的化合物和抗青光眼药物）是否对培养的视网膜神经节细胞（RGC）和视神经挤压（ONC）视网膜中的大鼠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。通过XTT试验和活化caspase-3的免疫细胞化学法，监测使用或不使用他氟前列素时的细胞死亡水平。用Fluo-4荧光测定细胞内钙（[Ca 2 +]i）水平的变化。ONC可使大鼠RGCs变性。他氟前列素局部滴注7天和14天后，对逆行标记的RGC数量进行计数。用末端dUTP缺口末端标记法（TUNEL）对视网膜平片进行染色以检测凋亡细胞。前列腺素剂量依赖性地促进RGC-5细胞的活力，最佳浓度为3 μ M（p = 0.006）。替前列素显著减少caspase-3阳性细胞，并抑制外源性谷氨酸引起的[Ca+2]i。cGMP依赖性蛋白激酶抑制剂和KT-5823部分阻断了他氟前列素的挽救作用（p = 0.002）。在接受他氟前列素治疗的眼睛中，RGCs的存活率显著增加（p = 0.01），ONC后14天TUNEL阳性细胞的发生率显著降低（p < 0.001）。这些数据表明，他氟前列素在RGCs中具有抗凋亡作用。
To investigate whether tafluprost, which is a prostaglandin-related compound and an anti-glaucoma drug, has a direct anti-apoptotic effect in cultured retinal ganglion cells (RGCs) and rat RGCs in retinas with optic nerve crush (ONC).RGC-5 cells were induced to undergo apoptosis by a serum deprivation and by exogenous glutamate. The level of cell death with or without tafluprost was monitored by an XTT assay and by immunocytochemistry with activated caspase-3. Changes in intracellular calcium ([Ca2+]i) levels were measured with fluo-4 fluorescence. Rat RGCs were degenerated by ONC. After topical instillation of tafluprost for 7 and 14 days, the numbers of retrograde-labeled RGCs were counted. Retinal flatmounts were subjected to terminal dUTP nick end labeling (TUNEL) staining to detect apoptotic cells.Tafluprost dose-dependently promoted RGC-5 cell viability with an optimum concentration of 3 mu M (p = 0.006). Tafluprost significantly reduced caspase-3-positive cells and suppressed [Ca+2]i evoked by exogenous glutamate. The cGMP-dependent protein kinase inhibitor and KT-5823 partially blocked the rescue effect of tafluprost (p = 0.002). The survival rate of RGCs significantly increased in eyes treated with tafluprost (p = 0.01), and the prevalence of TUNEL-positive cells was significantly decreased 14 days after ONC (p < 0.001).These data suggest that tafluprost has an anti-apoptotic effect in RGCs.