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目的 :观察睫状神经营养因子 (CNTF)对培养大鼠视网膜神经节细胞 (RGCs)凋亡的影响。方法 :15只生后 2～ 3dWistar大鼠 ,视网膜采用胰酶消化法制成细胞悬液后接种于 2 4孔培养板 (每孔 1× 10 5个细胞 ) ,取培养 72h的细胞行免疫细胞化学鉴定。将培养的细胞随机分为对照组、三种浓度CNTF组 (10ng/ml、2 0ng/ml、4 0ng/ml) ,培养 72h后采用TUNEL法和流式细胞仪检测培养大鼠视网膜神经节细胞凋亡的变化。结果 :培养 72h的细胞 90 %以上为视网膜神经节细胞 ,TUNEL法检测显示培养细胞有凋亡细胞存在 ,流式细胞仪检测结果显示对照组凋亡率为 (11.95± 4 .4 0 0 ) % ,10ng/ml、2 0ng/ml、4 0ng/mlCNTF组凋亡率分别为(7.883± 2 .14 6 ) % ,(6 .4 17± 3.317) % ,(8.0 33± 2 .0 2 3) % ,CNTF组凋亡率显著降低 (P <0 .0 1～ 0 .0 5 ) ,其中2 0ng/mlCNTF组差异非常显著 (P <0 .0 1)。结论 :CNTF能降低培养大鼠视网膜神经节细胞凋亡率 ,其促视网膜神经节细胞存活可能是通过减少视网膜神经节细胞凋亡来起作用 ,而高浓度CNTF对视网膜神经节细胞有毒副作用。
Objective: To observe the effect of ciliary neurotrophic factor (CNTF) on the apoptosis of cultured rat retinal ganglion cells (RGCs). METHODS: Fifteen postnatal 2 ~ 3d Wistar rats were made into retinas by trypsin digestion and seeded on 24-well plates (1 × 10 5 cells per well). The cells cultured for 72 hours were immunocytochemistry Identification. The cultured cells were randomly divided into control group, three concentrations of CNTF group (10ng / ml, 20ng / ml, 40ng / ml), cultured for 72h after TUNEL method and flow cytometry rat retinal ganglion cells Apoptosis changes. Results: More than 90% of cells cultured for 72 hours were retinal ganglion cells. TUNEL assay showed that there were apoptotic cells in the cultured cells. The results of flow cytometry showed that the apoptotic rate in the control group was (11.95 ± 4.40)% (7.883 ± 2.146)%, (6.417 ± 3.317)%, (8.033 ± 2.023)% respectively in 10ng / ml, 20ng / ml and 40ng / %, The apoptosis rate of CNTF group was significantly decreased (P <0.01 ~ 0.05), of which 20 ng / mlCNTF group was significantly different (P <0.01). CONCLUSION: CNTF can reduce the apoptosis rate of cultured rat retinal ganglion cells and promote the survival of retinal ganglion cells through reducing the apoptosis of retinal ganglion cells. CNTF with high concentration has toxic and side effects on retinal ganglion cells.