CAJ | 학술논문

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(25 mmol/L葡萄糖浓度)、高糖组(55 mmol/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100 mg/L、200 mg/L的黄芪甲苷组RGC-5细胞存活率分别81.6％、80.0％,与高糖组(66.0％)相比差异有统计学意义(P＜0.05),100 mg/L黄芪甲苷是其最佳保护浓度.100 mg/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.100 mg/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.
목적 탐토황기갑감대고당배양하시망막신경절세포( RGC)적보호작용.방법 장배양적RGC-5주분위대조조(25 mmol/L포도당농도)、고당조(55 mmol/L포도당농도)급황기갑감조,용세포기술시제합-8(CCK-8)측시망막신경절세포생존솔,이류식세포의평고황기갑감대고당배양적시망막신경절세포조망적영향,격광공취초급전경분별관찰각조선립체막전위급선립체형태변화.결과 불동농도황기갑감예처리균가제고RGC-5세포존활솔,기중100 mg/L、200 mg/L적황기갑감조RGC-5세포존활솔분별81.6％、80.0％,여고당조(66.0％)상비차이유통계학의의(P＜0.05),100 mg/L황기갑감시기최가보호농도.100 mg/L황기갑감예처리조RGC-5세포조망솔위6.1％,고당조적RGC-5세포적조망솔위8.2％(P＜0.05).전경관찰고당조RGC-5내포질수종,선립체종창,부분척소실,출현공포.100 mg/L황기갑감조,가견선립체결구청석완정,선립체경도수종,선립체척상가.황기갑감가상조선립체막전위,완해선립체수종.결론 황기갑감대고당인기적RGC억제유일정적보호작용.
Objective To explore the protective effects of astragaloside Ⅳ on retinal ganglion cells (RGC) incubated under high glucose.Methods Cultured 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.