睫状神经营养因子对培养大鼠视网膜神经节细胞的影响  

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

作　　者：袁容娣[1] 贺翔鸽[1] 叶剑[1] 彭锡嘉[1] 

机构地区：[1]第三军医大学大坪医院眼科,重庆400042

出　　处：《中华眼底病杂志》2002年第4期283-285,共3页Chinese Journal of Ocular Fundus Diseases

基　　金：国家基础研究发展规划 973资助项目 (1 9990 542 0 4 ) ;全军"十五"医药卫生科研基金资助项目 (0 1 MA1 75)

摘　　要：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、2 0、40 ng/ ml CNTF组 ( 、 、 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, 组 A值分别为 0 .0 75 8± 0 .0 13 9、0 .0 6 93± 0 .0 113, 组 A值分别为 0 .0 90 2± 0 .0 114、0 .0 82 5± 0 .0 12 5 , 组 A值分别为0 .0 792± 0 .0 13 3、0 .0 6 5 3± 0 .0 0 86 ,对照组 A值分别为 0 .0 6 2 0± 0 .0 0 71、0 .0 5 13± 0 .0 0 6 8。实验组与同时间对照组 A值相比差异有显著性的意义 ( 组与对照组相比 P<0 .0 1, 、 组与对照组相比 P<0 .0 5 )。　结论　一定浓度的 CNTF能促进培养大鼠 RGC的存活 ,?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: 0.075 8±0.013 9 and 0.069 3±0.011 3 in I group, 0.0902±0.011 4 and 0.082 5±0.0125 in Ⅱ group, 0.079 2±0.013 3 and 0.065 3±0.008 6 in Ⅲ group, and 0.062 0±0.007 1 and 0.051 3±0.006 8 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.

关 键 词：视网膜神经节细胞 细胞培养 睫状神经营养因子 CNTF 动物实验 免疫细胞化学 

分 类 号：R774[医药卫生—眼科]