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目的:研究N-甲基-D-天门冬氨酸(N-methyl-D-aspartate NMDA)及地塞米松对体外培养视网膜神经节细胞(RGCs)的作用。方法:出生1～3天Sprague-Dawley(SD)乳鼠做体外RGCs培养。NMDA按20～500μmol/L加入两组混合培养的RGCs中,24小时后一组用Hochest33258检测凋亡细胞。另一组加0.4%台盼蓝,计数拒染细胞,并算出细胞存活率。地塞米松按1×10~(-4)、1×10~(-5)及1×10~(-6)mol/L分别加入混合培养及纯化培养的RGCs中,对照组不加,24小时后用Hochest33258检测凋亡细胞。结果:混合培养RGCs经不同浓度NMDA处理均可见明显的凋亡细胞形态学改变。对照组则不明显。而且其中细胞的存活率与NMDA浓度呈反相关。不同浓度地塞米松处理混合培养RGCs,经Hochest33258检测均未发现有凋亡形态学改变。而纯化培养RGCs实验组及对照组均有明显凋亡细胞形态学改变。结论:NMDA对体外混合培养RGCs有诱导细胞凋亡的作用;地塞米松则对培养RGCs无诱导细胞凋亡作用。纯化的RGCs生长24小时,不加神经营养因子Neurotrophic Factors(NTFs)有细胞凋亡的形态学改变。眼科学报1999 15:65—69。
Objective: To investigate the effect of N-methyl-D-aspartate (NMDA) and dexamethasone on cultured retinal ganglion cells (RGCs) in vitro. Methods: Sprague-Dawley (SD) born 1 to 3 days old rabbits were cultured in vitro. NMDA was added into two groups of RGCs mixed with 20 ~ 500μmol / L, and 24 hours later, a group of apoptotic cells were detected by Hochest33258. The other group plus 0.4% trypan blue, counting of non-transfected cells, and calculate the cell viability. Dexamethasone was added into mixed culture and purified RGCs at 1 × 10 -4, 1 × 10 -5 and 1 × 10 -6 mol / L, respectively. The control group did not add 24 Hochest33258 was used to detect apoptotic cells after hours. Results: The morphological changes of apoptotic cells were observed after mixed culture of RGCs with different concentrations of NMDA. The control group is not obvious. Moreover, the cell survival rate was inversely correlated with the NMDA concentration. Different concentrations of dexamethasone mixed culture RGCs, Hochest33258 detected no apoptotic morphological changes. However, the morphological changes of apoptotic cells in purified RGCs group and control group were observed. CONCLUSION: NMDA induces apoptosis in RGCs cultured in vitro. Dexamethasone induces apoptosis in cultured RGCs. Purified RGCs grew for 24 hours without neurotrophic factors (NTFs) that had apoptotic morphological changes. Journal of Ophthalmology 1999 15: 65-69.