不同药物对视网膜神经细胞线粒体功能的影响研究

Effects of drugs on the functions of mitochondria in retinal nerve cells

目的 研究药物对视网膜神经细胞线粒体功能影响 ,为进行视网膜神经细胞药物保护研究提供依据。 方法 新生小牛 8只眼视网膜细胞培养 ,用激光扫描共聚焦显微镜 (Confocal)检测这些细胞在分别加入阿魏酸 (FA)、精氨酸、甘氨酸、牛磺酸、脑源性神经营养因子 (BDNF)和维生素 E前后 ,罗丹明12 3(dye rhodamine12 3,Rh12 3)标记的线粒体荧光强度的变化情况。 结果 5 0 0 μg/ ml FA可使线粒体荧光强度呈双相变化。扫描 6 0 .772 s首次给药后 ,荧光强度迅速下降 (4 5 .4 2 5± 4 .15 3→ 2 2 .135± 5 .2 93) ,但 112 .774 s再次给药后 ,曲线上扬 (19.6 5 5± 4 .383→ 2 8.2 4 7± 4 .76 4 ) ,在 16 8.773s第三次给药后 ,曲线仍上扬。 12 .5 mg/ ml维生素 E能增强线粒体荧光强度 ,扫描 5 6 .4 5 7s给药后 ,荧光强度曲线立即上扬(88.2 5 5± 5 .0 39→ 111.2 73± 4 .5 2 9)。 5 0 ng/ ml BDNF可使荧光强度升高 ,在 5 8.14 7、 134.14 8s两次给药后 ,荧光强度曲线均上扬 ,尤以首次作用明显 (6 9.115± 5 .0 38→ 77.2 2 5± 5 .131)。 2 .5 mg/ ml甘氨酸和30 mg/ ml精氨酸作用对线粒体荧光强度无明显影响。 6 .2 5 mg/ ml牛磺酸可使线粒体荧光强度轻度减弱。 结论 FA、BDNF、维生素 E可能通过线粒体途?

Objective To explore the effects of drugs on functions of mitochondria in retinal nerve cells, and to lay a foundation of the investigation of drug protection for retinal nerve cells. Methods Cultivation of the retinal nerve cells of 8 eyes of neonatal calves was performed. The changes of fluorescent density of the mitochondria of cultured cells labeled by dye rhodamine 123 (Rh123) before and after the activation of the medicines, including ferulic acid (FA), arginine, glycine, taurine, vitamine E and brain-derived neurotrophic factor (BDNF) respectively, were detected by laser-scanning confocal microscopy. Results FA with the concentration of 500 μg/ml led the diphasic variation of the fluorescent intensity of mitochondria. After scanning for 60.772 seconds when treated with FA firstly, the fluorescent intensity decreased rapidly (from 45.425±4.153 to 22.135±5.293); while after 112.774 seconds when treated secondly, the intensity increased obviously (from 19.655±4.383 to 28.247±4.764), and after 168.773 seconds when treated thirdly the intensity still increased. After scanning for 56.457 seconds when treated with vitamin E (12.5 mg/ml), the fluorescent intensity increased obviously (from 88.255±5.039 to 111.273±4.529), which suggested that vitamin E with the concentration of 12.5 mg/ml strengthen the fluorescent intensity. After scanning for 58.147 and 134.148 seconds when treated with BDNF (50 ng/ml) respectively, the fluorescent intensity increased obviously (from 69.115±5.038 to 77.225±5.131) which suggested that BDNF with the concentration of 50 ng/ml led the increase of the fluorescent intensity. Glycine (2.5 mg/ml) and arginine(30 mg/ml) didn't affect the fluorescent intensity of mitochondria, and taurine (6.25 mg/ml) caused the appreciable decrease of the fluorescent intensity. Conclusion FA, BDNF and vitamin E may promote the metabolism of retinal nerve cells via the path of mitochondria, while amino acids may adjust the activation of retinal nerve cells through other ways.