Latanoprost rescues retinal neuro-glial cells from apoptosis by inhibiting caspase-3, which is mediated by p44/p42 mitogen-activated protein kinase
Latanoprost rescues retinal neuro-glial cells from apoptosis by inhibiting caspase-3, which is mediated by p44/p42 mitogen-activated protein kinase
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DOI：
10.1016/j.exer.2006.05.018
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发表时间：
2006-11-01
影响因子：
3.4
通讯作者：
Negi, Akira
中科院分区：
文献类型：
--
作者：
Nakanishi, Yoriko;Nakamura, Makoto;Negi, Akira
本研究的目的是研究拉坦前列素（一种前列腺素F2 α类似物）是否在培养的视网膜神经胶质细胞和糖尿病视网膜中具有直接的抗凋亡作用。R28细胞是永生化的视网膜神经胶质祖细胞，24小时血清剥夺可诱导R28细胞凋亡。停药血清使15%的R28细胞发生固缩和活化的caspase-3免疫反应，拉坦前列素酸抑制凋亡，最佳浓度为1.0 μ M（P < 0.001），具有剂量依赖性。丝裂原活化蛋白激酶（MAPK）/细胞外信号调节激酶激酶（MEK）1和2抑制剂UO 126逆转了这种作用。链脲佐菌素诱导的1个月或3个月糖尿病大鼠左眼接受平衡盐溶液（BSS），右眼接受拉坦前列素滴眼液，持续5天。视网膜整体进行末端dUTP缺口末端标记（TUNEL）染色，而眼球摘除切割半胱天冬酶-3免疫荧光。检测视网膜匀浆中磷酸化或总p44/p42 MAPK和Akt。1个月和3个月的糖尿病视网膜每0.5 CM分别有30.2 +/- 15.3和23.6 +/- 9.0个TUNEL阳性细胞，而对照视网膜有很少的TUNEL阳性细胞。拉坦前列素滴注显著减少了这些细胞（1 M和3 M分别为10.0 +/- 3.1和11.3 +/- 3.1个细胞/0.5 cm（2），P < 0.01），而BSS没有。拉坦前列素还能显著减少神经节细胞和内核层的caspase-3免疫反应阳性细胞（P < 0.05）。拉坦前列素可增加p44/p42 MAPK磷酸化蛋白/总蛋白比值（P < 0.05），但对Akt无影响。总之，本研究结果表明，拉坦前列素拯救视网膜神经元和/或神经胶质细胞凋亡，这可能是通过抑制caspase-3介导的p44/p42 MAPK。(c)2006爱思唯尔有限公司保留所有权利。
The purpose of this study was to investigate whether latanoprost, a prostaglandin F2 alpha analogue, has a direct anti-apoptotic effect both in retinal neuro-glial cells in culture and in diabetic retina. R28 cells, immortalized retinal neuroglial progenitor cells, were induced apoptosis by 24 h serum deprivation. Serum withdrawal made up to 15% of R28 cells pyknotic and activated caspase-3 immunoreactive, and latanoprost acid suppressed apoptosis with dose dependency at an optimum concentration of 1.0 mu M (P < 0.001). UO126, a mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase kinase (MEK) 1 and 2 inhibitor reversed this effect. Streptozotocin induced one- or three-month diabetic rats received balanced-salt-solution (BSS) in the left eye and latanoprost eye drops in the right for 5 days. Retinal wholemount was subjected to terminal dUTP nick end labeling (TUNEL) staining, whereas eyeballs were enucleated for cleaved caspase-3 immunofluorescence. Retinal homogenates were probed for phospho- or total p44/p42 MAPK and Akt. One- and three-month diabetic retina had 30.2 +/- 15.3 and 23.6 +/- 9.0 TUNEL positive cells per 0.5 CM, respectively, whereas control retina had few TUNEL positive cells. Latanoprost instillation significantly reduced these cells (10.0 +/- 3.1 and 11.3 +/- 3.1 cells per 0.5 cm(2) for 1 M and 3 M, respectively, P < 0.01), whereas BSS did not. Latanoprost also significantly reduced cleaved caspase-3 immunoreactive cells in ganglion cell and inner nuclear layers (P < 0.05). Latanoprost increased phosphorylated to total protein ratio of p44/p42 MAPK (P < 0.05), but not of Akt. Taken together, the present findings suggest that latanoprost rescues retinal neurons and/or glial cells from apoptosis, which is probably mediated by p44/p42 MAPK through caspase-3 inhibition. (c) 2006 Elsevier Ltd. All rights reserved.