目的探讨黄芪甲苷对高糖培养下视网膜神经节细胞(RGC)的保护作用。方法将培养的RGC-5株分为对照组(25mmol/L葡萄糖浓度)、高糖组(55mmol/L葡萄糖浓度)及黄芪甲苷组,用细胞技术试剂盒-8(CCK.8)测视网膜神经节细胞生存率,以流式细胞仪评估黄芪甲苷对高糖培养的视网膜神经节细胞凋亡的影响,激光共聚焦及电镜分别观察各组线粒体膜电位及线粒体形态变化。结果不同浓度黄芪甲苷预处理均可提高RGC-5细胞存活率,其中100mg/L、200mg/L的黄芪甲苷组RGC-5细胞存活率分别81.6%、80.0%,与高糖组(66.O%)相比差异有统计学意义(P〈0.05),100mg/L黄芪甲苷是其最佳保护浓度。100mg/L黄芪甲苷预处理组RGC-5细胞凋亡率为6.1%,高糖组的RGC-5细胞的凋亡率为8.2%(P〈0.05)。电镜观察高糖组RGC.5内胞质水肿,线粒体肿胀,部分嵴消失,出现空泡。100mg/L黄芪甲苷组,可见线粒体结构清晰完整,线粒体轻度水肿,线粒体嵴尚可。黄芪甲苷可上调线粒体膜电位,缓解线粒体水肿。结论黄芪甲苷对高糖引起的RGC抑制有一定的保护作用。
Objective To explore the protective effects of astragaloside IV on retinal ganglion cells(RGC) incubated under high glucose. Methods Cuhured RGC-5 were divided into 3 groups: control, high glucose and high glucose + astragaloside IV. The survival rate of cell was measured by CCK-8 kit and flow cytometry. The changes of mitochondrial membrane potential were monitored by confocal laser scanning microscopy while those of RGC-5 mitochondria observed by electron microscopy. Results When the cell survival rate was 100% in the control group, the survival rate improved after 1 h 5, 10, 50, 100,200 μg/ml astragaloside IV pretreatment and high-glucose culture. The cell survival rates for 100 mg/L, 200 mg/L of astragaloside IV were 81.6%, 80. 0%, the difference from the high glucose group (66%) was significant ( P 〈 0.05 ) and 100 mg/L of astragaloside IV was the best protection concentration. The apoptotic rate for 100 mg/L astragaloside IV pretreated group of RGC-5 cell was 6. 1%. And it could partially inhibit the RGC-5 cell apoptosis caused by high glucose ( 8.2% ) ( P 〈 0.05 ). In the high-glucose group, the cells showed marked increases in mitochondrial swelling, especially for cristae. Numerous normal mitochondria were observed in 100 mg/L astragaloside IV pretreated group. Astragaloside IV could elevate mitochondrial membrane potential and alleviate mitochondrial swelling. Conclusion Astragaloside IV can protect RGC from high-glucose induced damage and may benefit the patients with diabetic retinopathy.