Dual action of nitric oxide on purely isolated retinal ganglion cells
Dual action of nitric oxide on purely isolated retinal ganglion cells
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DOI：
10.1076/ceyr.23.4.233.5454
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发表时间：
2001-01-01
影响因子：
2
通讯作者：
Tsukahara, S
中科院分区：
文献类型：
--
作者：
Kashiwagi, K;Iizuka, Y;Tsukahara, S
目的。探讨一氧化氮(NO)在视网膜神经节细胞(RGCs)存活中的作用。用两步淘洗法从出生后SD大鼠的视网膜上皮细胞中分离RGCs，并在化学定义的无血清培养液中培养。NO释放剂S-亚硝基-N-乙酰青霉胺(SNAP：500um，250um，100um，10um，1um，100 nm，10 nM)，NO清除剂，2-(4-carboxyphenyl)-4，4，5，5tetramethylimidazoline-1-oxyl-3-oxide钾盐(c-PTIO：100um，33um，10um，1um)，100um SNAP与33um c-PTIO，N(G)-硝基-L-精氨酸甲酯的混合物(L-名字：10 mm，5 mm，500 um，100 um或10 um)，或将他们的车辆加入纯RGC培养基中48小时。流式细胞仪检测大小视网膜节细胞的存活率。在100um时，SNAP以浓度依赖的方式显著降低RGC的存活。在小于或等于1um时，SNAP显著提高存活率，尤其是大型视网膜节细胞。C-PTIO和L-NAME可浓度依赖性地降低小鼠的存活率。100um SNAP和33um c-PTIO的混合物与单独加入时相比显著提高了RGC的存活率。这些结果表明，NO对RGCs具有神经保护和神经毒性作用，低浓度的NO对体外培养的新生RGCs的存活可能是有利的。
Purpose. The role of nitric oxide (NO) in the survival of retinal ganglion cells (RGCs) was investigated.Methods. RGCs were purely isolated from postnatal Sprague-Dawley rats by 2-step panning and were cultured in chemically defined serum free medium. An NO releaser, S-nitroso-N-acetylpenicillamine (SNAP: 500 muM, 250 muM, 100 muM, 10 muM, 1 muM, 100nM, and 10 nM), an NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5tetramethylimidazoline-1-oxyl-3-oxide potassium salt (c-PTIO: 100 muM, 33 muM, 10 muM, 1 muM), mixture of 100 muM SNAP and 33 muM c-PTIO, N(G)-nitro-L-arginine methyl ester (L-NAME: 10 mM, 5mM, 500 muM, 100 muM or 10 muM), or their vehicles were added to the medium of pure RGC culture for 48 hr. Survival rates of small and large RGCs were determined separately by flow cytometry.Results. At 100 muM, SNAP significantly reduced RGC survival in a concentration dependent manner. At less than or equal to1 muM, SNAP significantly increased survival, particularly of large RGCs. c-PTIO and L-NAME reduced the survival rates concentration-dependently. A mixture of 100 muM SNAP and 33 muM c-PTIO significantly improved RGC survival compared with when they were added on their own.Conclusions. These results indicate that NO exhibits neuroprotective and neurotoxic actions on RGCs and that low concentrations of NO may be beneficial for the survival of neonatal RGCs in vitro.