睫状神经营养因子和碱性成纤维细胞生长因子对视网膜神经细胞Bcl-2蛋白动态表达的影响

目的 探讨睫状神经营养因子(ciliary neurotrophic factor,CNTF)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对新生期小牛视网膜神经细胞中Bcl-2蛋白表达的影响.方法 以新生小牛视网膜为研究对象,传代培养视网膜神经细胞.培养细胞传至第2代,以加入CNTF和bFGF为加药组,正常对照组不加任何药物.在接种后第3天加药组分别加入终浓度为50 μg·L-1的CNTF和bFGF,检测给药后第2天、第4天、第6天,给药组内和正常对照组内不同时间点以及两组间对应时间点Bcl-2蛋白表达变化情况.结果 CNTF组Bcl-2蛋白的表达在给药后第2天、第4天、第6天相对灰度值分别为0.502±0.029、0.389±0.022、0.320±0.031,不同时间点组内比较差异均具有统计学意义(P值分别为0.006、0.002、0.035).bFGF组Bcl-2蛋白的表达在给药后第2天、第4天、第6天相对灰度值分别为0.481±0.034、0.568±0.068、0.615±0.083,不同时间点组内比较差异均无统计学意义(P值分别为0.171、0.143、0.521).正常对照组相应时间点相应灰度值分别为0.314±0.033、0.300±0.004、0.299±0.060,不同时间点组内比较差异均无统计学意义(P值分别为0.688、0.667、0.976).在给药后第2天、第4天,CNTF组Bcl-2蛋白的表达强度与正常对照组相比差异均具有统计学意义(P=0.002、P=0.002),第6天与相应对照组间差异无统计学意义(P =0.626).在给药后第2天、第4天、第6天,bFGF组Bcl-2蛋白的表达强度与正常对照组相比差异均具有统计学意义(P=0.004、P=0.011、P=0.029).结论 CNTF和bFGF能够促进体外培养的新生小牛视网膜神经细胞Bcl-2蛋白表达,提高细胞抗凋亡的能力.

Effects of CNTF and bFGF on dynamic expression of Bcl-2 protein in retinal nerve cells

Objective To explore the effects of ciliary neurotrophic factor (CNTF) and basic fibroblast growth factor (bFGF) on expression of bcl-2 protein in retinal nerve cells of neonatal calf.Methods Retinal cells suspension from neonatal calf was used.The 2nd generation of cultured retinal cells of neonatal calf was treated with CNTF (50 μg·L-1,CNTF group) or bFGF (50 μg·L-1,bFGF group) after sub-cultured 3 days.Those who treated without CNTF or bFGF were set as control group.The expression of bcl-2 protein in retinal nerve cells was analyzed after treated with CNTF or bFGF of 2 days,4 days and 6 days.Results In CNTF group,the relative gray value of bcl-2 protein expression at 2 days,4 days,6 days were 0.502±0.029,0.389±0.022,0.320±0.031,respectively,there were statistical differences among different time points(P=0.006,0.002,0.035).In bFGF group,the relative gray value of bcl-2 protein expression at 2 days,4 days,6 days were 0.481±0.034,0.568±0.068,0.615±0.083,respectively,there was no statistical difference among different time points (P=0.171,0.143,0.521).In control group,the relative gray value of bcl-2 protein expression at 2 days,4 days,6 days were 0.314±0.033,0.300±0.004,0.299±0.060,respectively,there was no statistical difference among different time points (P=0.688,0.667,0.976).Compared with control group,there were significant differences in bcl-2 protein expression in CNTF group at 2 days and 4 days(P=0.002,0.002),but no significant difference at 6 days(P=0.626).Compared with control group,there were significant differences in bcl-2 protein expression in bFGF group at 2 days,4 days and 6 days(P=0.004,0.011,0.029).Conclusion CNTF and bFGF can promote the expression of bcl-2 protein in retinal nerve cells and improve its ability of anti-apoptosis.