枸杞多糖对高糖所致视网膜神经节细胞凋亡、基因表达及延迟整流钾电流的影响

摘要

目的:研究枸杞多糖(LBP)对高糖所致视网膜神经节细胞凋亡、基因表达及延迟整流钾电流的影响.方法:培养RGC-5视网膜神经节细胞株并分为对照组、高糖组、LBP组,分别用普通DMEM培养基、高糖DMEM培养基以及含有500 ng/mL LBP的高糖DMEM培养基处理.处理后,采用Annexin V-FITC/PI试剂盒测定凋亡细胞数目,采用荧光定量PCR试剂盒测定凋亡基因及抗氧化基因的表达量,采用膜片钳检测延迟整流钾电流.结果:干预后12 h、24 h、36 h、48 h时,高糖组的细胞凋亡数目显著高于对照组,LBP组的细胞凋亡数目显著低于高糖组;干预后24 h、48 h时,高糖组细胞中c-fos、c-jun、caspase-3、caspase-9、Nrf-2、NQO1、HO-1的mRNA表达量以及IK、Gmax均显著高于对照组,V1/2显著低于对照组;LBP组细胞中c-fos、c-jun、caspase-3、caspase-9的mRNA表达量以及IK、Gmax均显著低于高糖组,LBP组细胞中Nrf-2、NQO1、HO-1的mRNA表达量以及V1/2均显著高于高糖组.结论:枸杞多糖能够减轻高糖所致视网膜神经节细胞凋亡并抑制延迟整流钾电流的幅度.%Objective: To study the effect of lycium barbarum polysaccharides (LBP) on high glucose-induced retinal ganglion cell apoptosis, gene expression and delayed rectifier potassium current.Methods: RGC-5 retinal ganglion cell lines were cultured and divided into control group, high glucose group and LBP group that were treated with normal DMEM, high-glucose DMEM as well as high-glucose DMEM containing 500 ng/mL LBP respectively.After treatment, the Annexin V-FITC/PI kits were used to measure the number of apoptotic cells, fluorescence quantitative PCR kits were used to determine the expression of apoptosis genes and antioxidant genes, and patch clamp was used to test delayed rectifier potassium current.Results: Twelve hours, 24 h, 36 h and 48 h after intervention, the number of apoptotic cells of high glucose group was significantly higher than that of control group, and the number of apoptotic cells of LBP group was significantly lower than that of high glucose group;24 h and 48 h after intervention, c-fos, c-jun, caspase-3, caspase-9, Nrf-2, NQO1 and HO-1 mRNA expression as well as IK and Gmax of high glucose group were significantly higher than those of control group while V1/2 was significantly lower than that of control group;c-fos, c-jun, caspase-3 and caspase-9 mRNA expression as well as IK and Gmax of LBP group were significantly lower than those of high glucose group, while Nrf-2, NQO1 and HO-1 mRNA expression as well as V1/2 of LBP group were significantly higher than those of high glucose group.Conclusions: LBP can reduce the high glucose-induced retinal ganglion cell apoptosis and inhibit the delayed rectifier potassium current amplitude.