睫状神经营养因子对培养大鼠视网膜神经节细胞的影响

目的 观察不同浓度睫状神经营养因子(ciliary neurotrophic factor，CNTF)对培养大鼠视网膜神经节细胞（retinal ganglion cell，RGC）生长、存活的影响。 方法 取15只生后2～3d Wistar大鼠视网膜组织进行细胞培养，通过Thy-1单克隆抗体免疫细胞化学对培养的RGC进行鉴定。实验分对照组和10、20、40 ng/mlCNTF组（Ⅰ、Ⅱ、Ⅲ组），记录RGC存活时间，将培养第3、5、7天的RGC行四甲基偶氮唑盐（methylthio tetrazole,MTT）法测量吸光度(A)值［旧称光密度(OD)］。 结果 Thy-1单克隆抗体免疫组织化学检查显示培养3d的存活细胞90%以上为RGC。细胞存活期间实验组与对照组细胞均无明显突起,细胞体积无明显增大，实验组细胞存活时间比对照组长3~4d。培养第5、7d，Ⅰ组A值分别为0.075 8±0.0139、0.0693±0.0113，Ⅱ组A值分别为0.0902±0.0114、0.0825±0.0125，Ⅲ组A值分别为0.0792±0.0133、0.0653±0.0086，对照组A值分别为0.0620±0.0071、0.0513±0.0068。实验组与同时间对照组A值相比差异有显著性的意义（Ⅱ组与对照组相比P＜0.01，Ⅰ、Ⅲ组与对照组相比P＜0.05）。 结论 一定浓度的CNTF能促进培养大鼠RGC的存活，对RGC形态无影响。 (中华眼底病杂志, 2002, 18: 283-285)

Effect of ciliary neurotrophic factor on rat''s retinal ganglion cell in vitro

Objective To observe the effect of ciliary neurotrophic factor (CNTF) with different concentrations on the growth and survival of rats’ retinal ganglion cells (RGC) in vitro. Methods The retinae of 15 Wistar rats which were 2 or 3 days after birth were dissociated into cell suspension with 0.05% trypsin digestion. After 3 days, cultured RGC were identified with immunohistochemistry method using anti-rat Thy-1.1 monoclonal antibody. Cultured RGC were divided into the 10, 20, 40 ng/ml CNTF group （Ⅰ,Ⅱ, and Ⅲgroup） and the control group respectively. The duration of living RGC was recorded. After 3, 5 and 7 days, the A value of living cells was tested by methylthio-tetrazole colorimetric microassay. Results The result of immunohistochemical examination showed that 90% of living cells cultured for 3 days were RGC. No protuberance or volume increase of RGC were observed in CNTF groups and the control group. The duration of the living RGC was prolonged 3 to 4 days in CNTF groups compared with the control group. The A values of living RGC at the 5th and 7th days in the CNTF groups and the control group were: 0.0758±0.0139 and 0.0693±0.0113 in I group, 0.0902±0.0114 and 0.0825±0.0125 in Ⅱ group, 0.0792±0.0133 and 0.0653±0.0086 in Ⅲ group, and 0.0620±0.0071 and 0.0513±0.0068 in the control group, respectively. The differences between the simultaneous CNTF and control group were significant (between Ⅱ group and the control group: P＜0.01; between Ⅰ and Ⅲ group, and the control group: P＜0.05). Conclusion CNTF with some certain concentrations could facilitate survival of RGC in vitro. CNTF has no effect on the conformation of RGC. (Chin J Ocul Fundus Dis, 2002, 18: 283-285)