论文部分内容阅读
目的:探讨低剂量哇巴因对B27剥夺诱发的螺旋神经节细胞损伤的保护作用,并初步探讨其保护机制。方法:螺旋神经节细胞培养第7天开始分组实验,设B27剥夺损伤组(损伤组)、B27剥夺加10nmol/L哇巴因保护组(保护组)和正常培养对照组(对照组)。各组继续培养48h后用FITC标记的Annexin-V和碘化丙啶双染试剂盒染色,用流式细胞仪检测各组螺旋神经节细胞损伤情况。各组螺旋神经节细胞分别于加药后6、12h两个时间点用免疫细胞化学方法检测细胞内Bcl-2表达水平。另外,应用螺旋神经节组织块分组培养48h后光学显微镜下观察轴(树)突生长情况。结果:流式细胞仪检测结果显示,保护组螺旋神经节细胞损伤率为(9.0±1.3)%,显著低于损伤组(32.8±2.4)%,与对照组(6.3±0.3)%相比无明显差异;免疫细胞化学结果显示,保护组螺旋神经节细胞6h点Bcl-2水平较损伤组和对照组增加。螺旋神经节组织块培养显示,保护组轴(树)突生长明显优于损伤组。结论:低浓度哇巴因对B27剥夺诱发的螺旋神经节细胞损伤具有保护作用,此作用可能通过上调螺旋神经节细胞内Bcl-2表达水平来实现。
Objective: To investigate the protective effect of low dose ouabain on spiral ganglion cell injury induced by B27 deprivation, and to explore its protective mechanism. Methods: Spiral ganglion cells were cultured on the 7th day. Group B27 deprivation injury (injury group), B27 deprivation plus 10nmol / L ouabain protection group (protective group) and normal culture control group (control group). Each group was cultured for 48h and then stained with FITC-labeled Annexin-V and propidium iodide double staining kit. The injury of spiral ganglion cells in each group was detected by flow cytometry. Spiral ganglion cells in each group were detected by immunocytochemistry at 6 and 12 h after dosing, respectively. In addition, the application of spiral ganglion tissue culture group 48h after the optical microscope under axonal growth observed. Results: The results of flow cytometry showed that the injury rate of spiral ganglion cells in protective group was (9.0 ± 1.3)%, which was significantly lower than that in injured group (32.8 ± 2.4)%, compared with that in control group (6.3 ± 0.3)% The results of immunocytochemistry showed that Bcl-2 level in protective group increased at 6h after injury compared with injury group and control group. Spiral ganglion tissue culture showed that the protective group axon (tree) sudden growth was significantly better than the injury group. CONCLUSION: Low concentration of ouabain has a protective effect on the injury of spiral ganglion cells induced by B27 deprivation, and this effect may be achieved by up-regulating the expression of Bcl-2 in spiral ganglion cells.