摘要
目的评价枸杞多糖对视网膜神经节细胞凋亡的保护作用。方法体外培养大鼠视网膜神经节细胞株RGC-5,采用谷氨酸作为细胞凋亡诱导剂建立RGC-5凋亡模型,终浓度为5 mmol·L-1的谷氨酸作用24 h可造成约50%的RGC-5凋亡,用此方法作为RGC-5凋亡模型。并用不同浓度(10 mg·L-1、20 mg·L-1、40 mg·L-1)枸杞多糖加入培养基干预24 h及48 h,使用四甲基偶氮唑盐(methyl thiazolyl tetrazolium,MTT)法和AnnexinV-FITC法检测细胞存活率及凋亡率。结果 MTT法检测结果显示:枸杞多糖作用24 h后,对照组存活率为48.7%,枸杞多糖组(10 mg·L-1、20 mg·L-1、40 mg·L-1)细胞存活率分别为53.1%、59.6%、66.9%,对照组与各浓度枸杞多糖组差异均有统计学意义(均为P<0.05);40 mg·L-1枸杞多糖组与20 mg·L-1枸杞多糖组之间差异有统计学意义(P<0.05)。枸杞多糖作用48 h,各浓度枸杞多糖组细胞存活率分别为60.5%、75.2%、88.6%,各浓度枸杞多糖组的RGC-5存活率明显高于对照组(均为P<0.05),且40 mg·L-1枸杞多糖组RGC-5存活率最高,与20 mg·L-1枸杞多糖组之间差异有统计学意义(P<0.05)。AnnexinV-FITC法检测结果显示:枸杞多糖作用24 h,对照组凋亡细胞(早期凋亡和晚期凋亡)所占比例为44%,枸杞多糖组(10 mg·L-1、20 mg·L-1、40 mg·L-1)凋亡细胞比例分别为20.6%、12.3%、8.5%,对照组与各浓度枸杞多糖组差异均有统计学意义(均为P<0.05);枸杞多糖作用48 h时,枸杞多糖组(10 mg·L-1、20 mg·L-1、40 mg·L-1)凋亡细胞比例分别为11.6%、6.2%、3.7%;24 h及48 h时,40 mg·L-1枸杞多糖组与20 mg·L-1枸杞多糖组之间凋亡细胞所占比例差异有统计学意义(P<0.05)。结论枸杞多糖对RGC-5具有一定的保护作用,并表现出一定程度的剂量-时间依赖性。
Objective To evaluate the anti-apoptosis effects of lycium barbarum polysaccharides （LBP）on cultured retinal ganglion cell （RGC）. Methods Rat RGC line RGC-5 were cultured in vitro, and apoptosis models of RGC-5 induced by glutamate was established. Then apoptosis models were cultured in different concentrations of LBP （ 10 mg · L^-1 ,20 mg·L^-1 ,40 mg · L^-1 ）for 24 hours and 48 hours,and MTT cell vi- ability assay and flow cytometry were used to calculate the survival and apoptotic ratio of RGC-5. Results MTT cell viability assay showed that the survival ratio after cul- tured with LBP for 24 hours in control group was 48.7% ,which in LBP group （ 10 mg·L^-1 ,20 mg·L^-1,40 mg·L^-1 ）were 53.1% ,59.6% and 66.9% ,respectively,there was statistical difference between 20 mg·L^-1 LBP group and 40 mg·L^-1 LBP group （P 〈 0. 05） ;The survival ratio after cultured with LBP for48 hours in LBP group （10 mg·L^-1 , 20 mg·L^-1 ,40 mg·L^-1）were 50.5% ,75.2% and 88.6% ,respectively,which were obvi- ously higher than that in control group,the 40 mg·L^-1 LBP group was the highest,there was statistical difference between 20 mg·L^-1 LBP group and 40 mg·L^-1 LBP group （P 〈 0. 05 ）. Flow cytometry showed that the apoptotic ratio （ early and middle apoptotic stages） of RGC-5 after cultured with LBP for 24 hours in control group was 44.0%, which in LBP group （10 mg mg·L^-1 ,20 mg mg·L^-1 ,40 mg mg·L^-1）were 20.6% ,12.3% and 8.5% ,re- spectively;The apoptotic ratio （ early and middle apoptotic stages） of RGC-5 after cul- tured with LBP for 48 hours in LBP group （ 10 mg mg·L^-1,20 mg·L^-1 ,40 mg·L^-1）were 11.6% ,6.2% and 3.7% ,respectively,there were statistical differences between 20 mg·L^-1 LBP group and 40 mg·L^-1 LBP group after cultured with LBP for 24 hours and 48 hours （ all P 〈 0. 05 ）. Conclusion LBP plays certain protective effects on RGC-5 with a dose dependent manner.
出处
《眼科新进展》
CAS
北大核心
2014年第2期119-122,共4页
Recent Advances in Ophthalmology
基金
山东中医药科技发展计划项目(编号:2011-130)~~
关键词
枸杞多糖
视网膜神经节细胞
细胞凋亡
lycium barbarum polysaccharides
retinal ganglion cell
apoptosis
