薯蓣皂苷元调节PINK1/Parkin通路对高糖诱导的视网膜血管内皮细胞损伤的影响OA
Effect of diosgenin on high glucose induced retinal endothelial cell damage by modulating PINK1/Parkin pathway
目的 探讨薯蓣皂苷元(DSG)调节PTEN诱导激酶1(PINK1)/Parkin通路对高葡萄糖(HG)诱导的人视网膜血管内皮(HRCE)细胞损伤的影响.方法 将HRCE细胞分为对照组、HG组、HG+DSG(100 μg/mL)组、HG+DSG+自噬抑制剂(Mdivi-1,25 μm)组.CCK-8法检测各组HRCE细胞增殖能力;流式细胞术检测细胞凋亡;透射电镜观察细胞自噬;荧光探针检测细胞中活性氧(ROS)活性;ELISA实验检测细胞上清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平以及细胞中超氧化物歧化酶(SOD)活性;Western blot实验检测HRCE细胞中PINK1/Parkin通路相关蛋白表达.结果 与对照组相比,HG组HRCE细胞中线粒体片段数量增加、自噬小体数量减少,细胞存活率、SOD活性、PINK1、Parkin和Beclin1蛋白表达降低(均P<0.05),细胞凋亡率、ROS活性、TNF-α和IL-6水平升高(均P<0.05);与HG组相比,HG+DSG组细胞中线粒体片段数量减少,自噬小体、自噬溶酶体数量增加,细胞存活率、SOD活性、PINK1、Parkin和Beclin1蛋白表达升高(均P<0.05),细胞凋亡率、ROS活性、TNF-α和IL-6水平降低(均P<0.05);与HG+DSG组相比,HG+DSG+Mdivi-1组细胞中线粒体片段数量增加,自噬小体数量减少,细胞存活率、SOD活性、PINK1、Parkin和Beclin1蛋白表达降低(均P<0.05),细胞凋亡率、ROS活性、TNF-α和IL-6水平升高(均P<0.05).结论 DSG可能通过调控HG诱导的HRCE细胞中PINK1/Parkin通路蛋白表达,促进HRCE细胞增殖和自噬,抑制细胞凋亡,降低炎性因子分泌,缓解细胞氧化应激损伤.
Objective To investigate the effect of diosgenin(DSG)on high glucose(HG)-induced damage to hu-man retinal capillary endothelial cells(HRCE)by modulating the PTEN-induced kinase 1(PINK1)/Parkin pathway.Methods HRCE cells were assigned to the control group,HG group,HG+DSG(100 μg/mL)group,and HG+DSG+au-tophagy inhibitor(Mdivi-1,25 μm)group.The CCK-8 method was applied to test the proliferation ability of HRCE cells.Flow cytometry was performed to test cell apoptosis.Transmission electron microscopy was applied to observe cellular au-tophagy.Fluorescent probe was performed to test reactive oxygen species(ROS)activity.Enzyme-linked immunosorbent as-say(ELISA)experiment was used to measure tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the cell super-natant,and the activity of superoxide dismutase(SOD).Western blot experiments were used to detect PINK1/Parkin path-way related proteins in HRCE cells.Results Compared to the control group,the HG group showed an increase in the num-ber of mitochondrial fragments and a decrease in the number of autophagosomes in HRCE cells,and the cell survival rate,SOD activity,PINK1,Parkin,and Beclin1 proteins reduced manifestly(all P<0.05),while cell apoptosis rate,ROS ac-tivity,TNF-α and IL-6 levels increased clearly(all P<0.05).Compared to the HG group,the HG+DSG group showed a decrease in the number of mitochondrial fragments and an increase in the number of autophagosomes and autolysosomes,and the cell survival rate,SOD activity,PINK1,Parkin,and Beclin1 proteins increased clearly(all P<0.05),while cell apoptosis rate,ROS activity,TNF-α,and IL-6 levels reduced manifestly(all P<0.05).Compared to the HG+DSG group,the HG+DSG+Mdivi-1 group showed an increase in the number of mitochondrial fragments in cells and a decrease in the number of autophagosomes,the cell survival rate,SOD activity,PINK1,Parkin,and Beclin1 proteins reduced manifestly(all P<0.05),while cell apoptosis rate,ROS activity,TNF-α,and IL-6 increased clearly(all P<0.05).Conclusions DSG may promote proliferation and autophagy of HRCE cells,inhibit apoptosis,reduce inflammatory cytokine secretion,and alleviate oxidative stress damage by modulating PINK1/Parkin pathway proteins induced by HG.
都艳红;张士宏;夏春晓;王艳新
072750 河北保定,保定市第二中心医院眼科072750 河北保定,保定市第二中心医院眼科072750 河北保定,保定市第二中心医院眼科072750 河北保定,保定市第二中心医院眼科
薯蓣皂苷元PTEN诱导激酶1/Parkin通路视网膜血管内皮细胞
DiosgeninPTEN-induced kinase 1/Parkin pathwayHuman retinal endothelial cells
《临床眼科杂志》 2026 (2)
104-109,6
河北省保定市科技计划项目(2141ZF218)
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