铁皮石斛多糖对缺氧/复氧视网膜神经节细胞损伤的影响  

Effect of polysaccharides of dendrobium candidum on retinal ganglion cells damage due to hypoxia/reoxygenation

作　　者：肖杨晶 陈婷[1] XIAO Yangjing;CHEN Ting(The First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第一医院,哈尔滨150001

出　　处：《中国中医眼科杂志》2020年第9期613-617,共5页China Journal of Chinese Ophthalmology

摘　　要：目的探讨铁皮石斛多糖(PDC)对缺氧/复氧损伤视网膜神经节细胞RGC-5的保护作用,并探讨其机制。方法培养RGC-5细胞,造成缺氧/复氧损伤模型,分为对照组、模型组、PDC低浓度组及PDC高浓度组,MTT法观察细胞活性,测各组细胞上清液乳酸脱氢酶(LDH)及细胞超氧化物歧化酶(SOD)、丙二醛(MDA)的变化,流式细胞技术检测RGC-5细胞内活性氧表达,Hoechst33258染色检测凋亡细胞比例,Western blot分析bcl-2及Bax抗体的变化。结果(1)细胞存活率:模型组较对照组降低(t=19.620,P=0.000),差异有统计学意义。PDC低浓度组(t=4.518,,P=0.002)和PDC高浓度组(t=6.399,P=0.000)均高于模型组,差异具有统计学意义。(2)LDH释放量:模型组较对照组增加(t=25.576,P=0.000),差异有统计学意义。PDC低浓度组(t=13.396,P=0.000)及PDC高浓度组(t=18.446,P=0.000)均低于模型组,差异有统计学意义。(3)SOD水平:模型组较对照组降低(t=21.198,P=0.000),差异有统计学意义;PDC低浓度组(t=7.981,P=0.000)及PDC高浓度组(t=12.007,P=0.000)均高于模型组,差异具有统计学意义。(4)MDA水平:模型组较对照组升高(t=7.461,P=0.000),差异有统计学意义;PDC低浓度组(t=2.865,P=0.021)及PDC高浓度组(t=4.668,P=0.002)均低于模型组,差异具有统计学意义。(5)ROS的表达:对照组ROS主峰位于基线左侧,模型组细胞内ROS表达升高,主峰右移,PDC作用于RGC-5细胞后,细胞内ROS表达下降,ROS主峰逐渐左移。(6)RGC-5细胞凋亡:对照组未见明显凋亡细胞,细胞核均为暗蓝色低荧光;模型组细胞核着色不均匀,凋亡细胞呈亮蓝色高荧光;PDC处理后亮蓝色的凋亡细胞数量逐渐减少。(7)Bcl-2及Bax蛋白表达水平:模型组Bcl-2蛋白相对表达量较对照组降低(t=46.500,P=0.000),差异有统计学意义。PDC低浓度组(t=6.228,P=0.000)和PDC高浓度组(t=6.871,P=0.000)均高于模型组,差异具有统计学意义。模型组Bax蛋白相对表达量较对照组升高(t=7.766,P=0.000),差�OBJECTIVE To investigate the protective effect and mechanism of polysaccharides of dendrobium candidum(PDC)on retinal ganglion cells damage due to hypoxia/reoxygenation.METHODS RGC-5 cells were cultured and were induced into hypoxia/reoxygenation models,then divided into control group,model group,low concentration PDC group and high concentration PDC group.The cell survival rate was detected by MTT colorimetry,the changes of lactate dehydrogenase(LDH),superoxide dismutase(SOD)and malondialdehyde(MDA)in cell supernatant were measured,ROS expressions inside the cells were detected by flow cytometry technology,apoptotic cells proportion were detected by Hoechst33258 staining,the expressions of Bcl-2 and Bax antibody changes were tested by western blot.RESULTS(1)Cell survival rate:the survival rate of RGC-5 cells in model group was lower than that in control group,the difference was statistically significant(t=19.620,P=0.000).The survival rates of RGC-5 cells in low concentration PDC group(t=4.518,P=0.002),and high concentration PDC group(t=6.399,P=0.000)were higher than those in model group,the difference was statistically significant.(2)LDH release:the release of LDH in extracellular medium of model groupwas higher than that of control group,and the difference was statistically significant(t=25.576,P=0.000).Low concentration PDC group(t=13.396,P=0.000)and high concentration PDC group(t=18.446,P=0.000)were lower than those in model group,the difference was statistically significant.(3)SOD level:the activity of SOD in model groupwas lower than that in control group,and the difference was statistically significant(t=21.198,P=0.000);the activity of SOD in low concentration PDC group(t=7.981,P=0.000)and high concentration PDC group(t=12.007,P=0.000)was higher than that in model group,the difference was statistically significant.(4)MDA level:MDA level in model group was higher than it in control group,and the difference was statistically significant(t=7.461,P=0.000);MDA level in low concentration PDC group(t=2.865,P=0.02

关 键 词：铁皮石斛多糖 缺氧/复氧 视网膜神经节细胞 凋亡 

分 类 号：R774.1[医药卫生—眼科]