脑源性神经营养因子对视网膜神经细胞bcl-2蛋白动态表达的影响

Brain-derived neurotrophic factor on dynamic expression of bcl-2 protein in retinal nerve cells

目的探讨脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)对新生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BDNF为BDNF组,不加任何药物为正常对照组。在接种后第3天BDNF组加入终浓度为50μg.L-1的BDNF,Western blotting法检测给药后第2天、第4天、第6天BDNF组和正常对照组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BDNF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0.447±0.052、0.355±0.039、0.287±0.031,呈现逐渐下降的趋势,第2天与第4天(t=2.788,P=0.016)、第2天与第6天(t=4.844,P=0.000)比较差异均有统计学意义,第4天与第6天(t=2.056,P=0.062)比较差异无统计学意义。而正常对照组分别为0.314±0.033、0.300±0.004、0.299±0.060,第2天与第4天(t=0.411,P=0.688)、第2天与第6天(t=0.441,P=0.667)、第4天与第6天(t=0.030,P=0.976)之间比较差异均无统计学意义。在给药后第2天,BDNF组bcl-2蛋白的表达强度与正常对照组相比差异具有统计学意义(t=4.022,P=0.002),第4天(t=1.645,P=0.126)、第6天(t=0.381,P=0.710)与相应对照组间比较差异均无统计学意义。结论BDNF能在短期内促进视网膜神经细胞bcl-2蛋白表达,提高细胞抗凋亡的能力,对延缓退行性视网膜疾病中的凋亡形成,开展视网膜神经细胞的神经保护具有一定作用。

Objective To explore the effects of brain-derived neurotrophic factor（BDNF） on expression of bcl-2 protein in retinal nerve cells of newborn calf.Methods Retinal nerve cells of newborn calf were subcultured.As cultured to the second passage,the cells with BDNF were treated as BDNF group,without any drug treatment as normal control group.BDNF group was treated with 50 μg·L-1 BDNF at the 3rd day after inoculation.The expression changes of bcl-2 protein were detected at the 2nd day,the 4th day and the 6th day after drug treatment in BDNF group and normal control group and at corresponding time points between two groups with Western blotting.Results In BDNF group,the relative grey values of bcl-2 protein expression were 0.447±0.052,0.355±0.039 and 0.287±0.031 at the 2nd day,the 4th day and the 6th day after drug treatment,respectively,showing a descending trend.There was significant difference in relative grey value between at the 2nd day and the 4th day after drug treatment（t=2.788,P=0.016） and between at the 2nd day and the 6th day after drug treatment（t=4.844,P=0.000）,but no significant difference between at the 4th day and the 6th day after drug treatment（t=2.056,P=0.062）.In normal control group,the relative grey values were 0.314±0.033,0.300±0.004 and 0.299±0.060 at the 2nd day,the 4th day and the 6th day after drug treatment,respectively.There was no significant difference in relative grey value between at the 2nd day and the 4th day after drug treatment（t=0.411,P=0.688）,between at the 2nd day and the 6th day after drug treatment（t=0.441,P=0.667） or between at the 4th day and the 6th day after drug treatment（t=0.030,P=0.976）.There was statistical difference in expression of bcl-2 protein between BDNF group and normal control group at the 2nd day after drug treatment（t=4.022,P=0.002）,but no significant difference was found at the 4th day after drug treatment between two groups（t=1.645,P=0.126） or at the 6th day after drug treatment between two groups（t=0.381,P=0.710）.Conclusions BDNF can promote the expression of bcl-2 protein in retinal nerve cells in a short time and advance the ability of anti-apoptosis of retinal nerve cells.It has neuroprotective effects on retinal nerve cells in delaying apoptosis of retrogressive retinopathy.