天麻素对高糖培养的小胶质细胞和视网膜神经节细胞相互作用的影响  

Effect of Gastrodin on Interactions Between Microglia and Retinal Ganglion Cells Cultured by High Glucose

作　　者：李红梅[1,2,3] 雷霍[1,2,3] 杨晓春[1,2,3] 王瑞丽[1,2,3] 梅妍[1,2,3] 倪宁华[1,2,3] 赵海燕[1,2,3] 陈臻[1,2,3] 

机构地区：[1]昆明理工大学附属医院,云南昆明650032 [2]云南省第一人民医院眼科,云南昆明650032 [3]云南省眼底病研究中心,云南昆明650032

出　　处：《昆明理工大学学报（自然科学版）》2015年第6期96-102,共7页Journal of Kunming University of Science and Technology(Natural Science)

基　　金：云南省应用基础研究计划项目(2014FB093);云南省卫计委内设研究机构科技计划项目(2014NS287);王陇德院士工作站专项经费

摘　　要：将原代培养的小鼠视网膜神经节细胞（Retinal Ganglion Cells,RGCs）和BV2小鼠小胶质细胞株分别培养并传代,取3~5代进行实验.高糖条件下共培养两种细胞,并给予不同浓度天麻素溶液干预.倒置显微镜下观察两种细胞的形态及共生关系并拍照,Western blotting检测各组上清液神经营养因子前体蛋白（Pro-Nerve Growth Factor,pro NGF）的表达,并进行统计学分析.结果显示高糖组RGCs较对照组增生减弱、轴突减少,而BV2细胞增生则较对照组明显活跃,两种细胞聚集减少;加入天麻素后,高糖培养的RGCs增生较高糖组活跃,轴突增长,BV2细胞减少,两种细胞聚集增加.这种变化与天麻素的浓度成正比.Western blotting条带及半定量分析显示,对照组pro NGF的表达较低,高糖组明显升高,低、中浓度天麻素组pro NGF的表达与高糖组没有明显差异,但高浓度天麻素组pro NGF的表达则较高糖组明显降低.因此天麻素能改善高糖对RGCs形态和与小胶质细胞聚集作用的影响,并能降低pro NGF的表达,这可能是天麻素对糖尿病早期RGCs损伤的保护机制之一.Primary culture retinal ganglion cells（ RGCs） and BV2 mouse Microglia are cultured and passaged,3- 5 generations from which are taken to experiments. Two kinds of cells are cultured together in high glucose and given different concentration of gastrodin. The morphology of the two kinds of cells and symbiotic relationship between the two cells are observed and photographed under an inverted microscope. Western blotting is adopted to detect the expression of pro- nerve growth factor（ pro NGF） in cell supernatant from each group and statistical analysis is carried out. The results show that hyperplasia of RGCs in high glucose group is weakened and axons are reduced compared with those of the control group,while the BV2 cells proliferation is more active and aggregation of the two cells is reduced compared with those of the control group. After treatment with gastrodin,proliferation of RGCs in high glucose is more active,axons grow longer,and quantity of BV2 cells is reduced,meanwhile aggregation of both kinds of cells is increased compared with those of high glucose group. This change is dependent on the concentration of gastrodin. Western blotting strips and semi- quantitative analysis show that the expression of pro NGF in the control group is lower while that of the high glucose group is significantly higher,and there is no significant difference of pro NGF expression in the low and moderate concentration gastrodin groups compared with that of high glucose group,but the expression of pro NGF in the high concentration gastrodin group is significantly reduced compared with that of the high glucose group. Gastrodin is proved to improve the effects of high glucose on RGCs morphology and aggregation with microglial cells and to decrease the expression of pro NGF,which may be one of the protective mechanisms of gastrodin against RGCs damage in early diabetes.

关 键 词：天麻素 神经生长因子 糖尿病视网膜病变 小胶质细胞 视网膜神经节细胞 

分 类 号：R774.13[医药卫生—眼科]