桃叶珊瑚苷调节Ras/Raf/MEK/ERK通路对高糖诱导的视网膜神经节细胞炎性损伤的影响OA
Effect of aucubin on high glucose-induced inflammatory injury in retinal gan-glion cells by regulating the Ras/Raf/MEK/ERK pathway
目的 探讨桃叶珊瑚苷(AU)调节Ras/Raf/MEK/ERK通路对高糖诱导的视网膜神经节细胞(RGCs)炎性损伤的影响.方法 RGCs分为空白组(5.5 mmol·L-1葡萄糖培养)、诱导组(30.0 mmol·L-1 葡萄糖培养)、AU-L 组(30.0 mmol·L-1 葡萄糖+50 μmol·L-1AU 培养)、AU-H 组(30.0 mmol·L-1 葡 萄糖+100 μmol·L-1 AU 培养)、RasAL1 组(30.0 mmol·L-1 葡萄糖+2 μg·L-1Ras 激活剂 RasAL1 培养)以及 AU-H+RasAL1 组(30.0 mmol·L-1 葡萄糖+100 μmol·L-1 AU+2 μg·L-1 RasAL1 培养),均培养 48 h.CCK-8检测细胞增殖水平变化;CalceinAM/PI双染色法及流式细胞术检测细胞凋亡率;相关试剂盒检测细胞中活性氧(ROS)、谷胱甘肽过氧化物酶(GSH-PX)以及丙二醛(MDA)表达水平;qRT-PCR检测细胞白细胞介素(IL)-6、IL-1β以及肿瘤坏死因子(TNF-α)的mRNA表达水平;Western blot检测细胞Ras/Raf/MEK/ERK通路相关蛋白表达水平.结果 与空白组相比,诱导组细胞增殖水平与GSH-PX表达水平均降低,细胞凋亡率、ROS与MDA表达水平、IL-6、IL-1β以及 TNF-α mRNA表达水平、MEK、Ras、Raf以及ERK磷酸化表达水平均增加;与诱导组相比,RasAL1组细胞增殖水平与GSH-PX表达水平均降低,细胞凋亡率、ROS与MDA表达水平、IL-6、IL-1β以及TNF-α mRNA表达水平、MEK、Ras、Raf以及ERK磷酸化表达水平均增加,而AU-L组与AU-H组细胞增殖水平与GSH-PX表达水平均增加,细胞凋亡率、ROS与MDA表达水平、IL-6、IL-1β以及TNF-α mRNA表达水平、MEK、Ras、Raf以及ERK磷酸化表达水平均降低;与AU-H组相比,AU-H+Ras-AL1 组细胞增殖水平与GSH-PX表达水平均降低,细胞凋亡率、ROS与MDA表达水平、IL-6、IL-1β以及TNF-α mRNA表达水平、MEK、Ras、Raf以及ERK磷酸化表达水平均增加;与RasAL1组相比,AU-H+RasAL1组细胞增殖水平与GSH-PX表达水平均增加,细胞凋亡率、ROS与MDA表达水平、IL-6、IL-1β以及 TNF-α mRNA表达水平、MEK、Ras、Raf以及ERK磷酸化表达水平均降低,差异均有统计学意义(均为P＜0.05).结论 AU通过抑制Ras/Raf/MEK/ERK通路减轻高糖诱导的RGCs炎性损伤.
Objective To investigate the effect of aucubin(AU)on high glucose-induced inflammatory injury in reti-nal ganglion cells(RGCs)by modulating the Ras/Raf/MEK/ERK pathway.Methods RGCs were divided into the blank group(cultured with 5.5 mmol·L-1 glucose),the induction group(cultured with 30.0 mmol·L-1 glucose),the AU-L group(cultured with 30.0 mmol·L-1 glucose and 50 μmol·L-1 AU),the AU-H group(cultured with 30.0 mmol·L-1 glu-cose and 100 μmol·L-1 AU),the RasAL1 group(cultured with 30.0 mmol·L-1 glucose and 2 μg·L-1 Ras activator RasAL1),and the AU-H+RasAL1 group(cultured with 30.0 mmol·L-1 glucose,100 μmol·L-1 AU,and 2 μg·L-1 RasAL1).All groups were cultured for 48 h.Cell proliferation was assessed by CCK-8 assay;apoptosis rate was evaluated by CalceinAM/PI double staining and flow cytometry;the expression levels of reactive oxygen species(ROS),glutathione per-oxidase(GSH-PX),and malondialdehyde(MDA)were measured using corresponding assay kits;mRNA expression levels of interleukin(IL)-6,IL-1β,and tumor necrosis factor(TNF)-α were detected by qRT-PCR;and the expression levels of Ras/Raf/MEK/ERK pathway-related proteins were examined by Western blot.Results Compared with the blank group,the in-duction group exhibited decreased cell proliferation and GSH-PX level,increased apoptosis rate,elevated ROS and MDA lev-els,upregulated mRNA expression of IL-6,IL-1β,and TNF-α,and enhanced phosphorylation of MEK,Ras,Raf,and ERK.Compared with the induction group,the RasAL1 group showed decreased cell proliferation and GSH-PX level,increased ap-optosis rate,elevated ROS and MDA levels,upregulated mRNA expression of IL-6,IL-1β,and TNF-α,and enhanced phospho-rylation of MEK,Ras,Raf,and ERK,whereas both the AU-L and AU-H groups exhibited increased cell proliferation and GSH-PX level,decreased apoptosis rate,reduced ROS and MDA levels,downregulated mRNA expression of IL-6,IL-1 β,and TNF-α,and diminished phosphorylation of MEK,Ras,Raf,and ERK.Compared with the AU-H group,the AU-H+RasAL1 group showed decreased cell proliferation and GSH-PX level,increased apoptosis rate,elevated ROS and MDA levels,upregulated mRNA expression of IL-6,IL-1β,and TNF-α,and enhanced phosphorylation of MEK,Ras,Raf,and ERK.Compared with the RasAL1 group,the AU-H+RasAL1 group exhibited increased cell proliferation and GSH-PX level,decreased apoptosis rate,reduced ROS and MDA levels,downregulated mRNA expression of IL-6,IL-1β,and TNF-α,and diminished phosphorylation of MEK,Ras,Raf,and ERK.All differences were statistically significant(all P＜0.05).Conclusion AU attenuates high glucose-induced inflammatory injury in RGCs by inhibiting the Ras/Raf/MEK/ERK pathway.
康健;郑雪;曹朗
072750 河北省保定市,保定市第二中心医院眼科072750 河北省保定市,涿州市医院神经内科072750 河北省保定市,保定市第二中心医院眼科
医药卫生
桃叶珊瑚苷高糖视网膜神经节细胞Ras/Raf/MEK/ERK通路
aucubinhigh glucoseretinal ganglion cellsRas/Raf/MEK/ERK pathway
《眼科新进展》 2026 (9)
694-699,6
保定市科技计划项目(编号:2141ZF218)
评论