人参皂苷Rg1对缺氧诱导所致人视网膜色素上皮细胞损伤的抑制作用及机制

Inhibitory effect of ginsenoside Rg1 on hypoxia-induced human retinal pigment epithelial cell injury and its mechanism

目的探究人参皂苷Rg1(GRg1)对缺氧诱导所致人视网膜色素上皮(ARPE)细胞损伤的作用及机制。方法体外培养ARPE-19细胞,将其随机分为对照组、模型组及GRg1高、中、低剂量组与缺氧诱导因子1α(HIF-1α)抑制剂组。除对照组外,其余组采取氯化钴诱导ARPE-19细胞构建缺氧模型,通过CCK-8法、TUNEL法测定各组不同时间点细胞活性及凋亡情况,DCFH-DA荧光探针对各组活性氧(ROS)水平,经由RT-qPCR、Western blotting法测定各组细胞内HIF-1α、BDNF、PACAP38 mRNA与蛋白表达。结果与对照组相比,各组给药0、12、24 h时ARPE-19细胞活性均低、凋亡率高(P均<0.05);GRg1高剂量组给药12、24、48 h时ARPE-19细胞活性均高于模型组、GRg1低剂量组(P均<0.05),细胞凋亡率均低于模型组、GRg1低剂量组(P均<0.05);GRg1高剂量组给药24、48 h时细胞活性均高于GRg1中剂量组(P均<0.05)。与对照组相比,各组给药48 h时ARPE-19细胞ROS、HIF-1αmRNA及蛋白表达均高(P均<0.05);GRg1高剂量组ARPE-19细胞ROSHIF-1αmRNA及蛋白表达均低于模型组、GRg1低剂量组(P均<0.05);GRg1高剂量组ARPE-19细胞BDNF、PACAP38 mRNA及蛋白表达均高于模型组、GRg1低剂量组(P均<0.05);GRg1高剂量组与HIF-1α抑制剂组给药不同时间点细胞活性与凋亡率、细胞ROS、HIF-1α、BDNF、PACAP38表达比较差异均无统计学意义(P均>0.05)。结论GRg1能减轻缺氧诱导所致ARPE-19细胞损伤,其机制可能与抑制细胞ROS、HIF-1α表达,上调BDNF、PACAP38表达有关。

Objective To explore the effect and mechanism of ginsenoside Rg1(GRg1)on hypoxia-induced human retinal pigment epithelium(RPE)cell injury.Methods ARPE-19 cells were cultured in vitro and randomly divided into control group,model group,high-dose GRg1 group,medium-dose GRg1 group,low-dose GRg1 group,and hypoxia-induced factor 1α(HIF-1α)inhibitor group,respectively.Except for the control group,ARPE-19 cells in the other groups were induced by cobalt chloride to construct anoxic model.CCK-8 method and TUNEL method were used to determine cell activity and apoptosis in each group at different time points.The levels of reactive oxygen species(ROS)in each group were measured by DCFH-DA fluorescent probe,and HIF-1α,brain-derived neurotrophic factor(BDNF)and pituitary adenylate cyclase activating polypeptide(PACAP)38 mRNA and protein expression levels in each group were measured by RT-qPCR and Western blotting.Results Compared with the control group,the other groups had significantly lower ARPE-19 cell activity and significantly higher apoptosis rates at 0,12 and 24 h after administration(all P<0.05).At 12,24 and 48 h after administration,the activity of ARPE-19 cells in the high-dose GRg1 group was obviously higher than that in the model group and low-dose GRg1 group(both P<0.05),while the apoptosis rate was obviously lower than those in the model group and low-dose GRg1 group(both P<0.05);the cell activity in the high-dose GRg1 group was significantly higher than that in the medium-dose GRg1 group at 24 and 48 h after administration(both P<0.05).Compared with the control group,the other groups had significantly higher ROS and HIF-1αmRNA and protein expression levels in ARPE-19 cells at 48 h after administration(all P<0.05).In the high-dose GRg1 group,the expression levels of ROSHIF-1αmRNA and protein in ARPE-19 cells were obviously lower than those in the model group and low-dose GRg1 group(all P<0.05),while BDNF,PACAP38 mRNA and protein expression levels in ARPE-19 cells were significantly higher than those in the model group and low-dose GRg1 group(all P<0.05).There were no statistically significant differences in cell activity,apoptosis rate,ROS,HIF-1α,BDNF or PACAP38 expression levels between high-dose GRg1 group and HIF-1αinhibitor group at different time points(all P>0.05).Conclusion GRg1 can alleviate the hypoxia-induced ARPE-19 cell injury,and its mechanism may be related to inhibiting the expression levels of ROS and HIF-1α,and up-regulating the expression levels of BDNF and PACAP38.