葡萄籽原花青素通过Xc^(-)/GPX4通路拮抗高糖高脂诱导的MIN6细胞铁死亡

Grape seed proanthocyanidins exacts antagonize high-glucose and high-fat-induced ferroptosis in MIN6 cells via the Xc^(-)/GPX4 pathway

目的探讨葡萄籽原花青素(GSPE)对高脂高糖诱导小鼠胰岛素瘤MIN6细胞损伤的保护作用及可能机制。方法采用CCK-8法检测细胞活力;Lillie染色检测细胞内亚铁离子(Fe^(2+));检测谷胱甘肽(GSH),丙二醛(MDA)和超氧化物歧化酶(SOD)含量;荧光探针DCFH-DA标记检测活性氧簇(ROS)水平;Western blot检测铁死亡关键蛋白(GPX4和ACSL4)和Xc^(-)/GPX4信号通路蛋白(SLC7A11、SLC3A2、GSS和GCLM)的表达。结果高脂高糖处理抑制了MIN6细胞活力,使得Fe^(2+),MDA含量,ROS水平增加,GSH,SOD含量降低,ACSL4表达水平升高,GPX4以及通路蛋白SLC7A11、SLC3A2、GSS和GCLM表达水平降低。施加不同浓度的GSPE或铁死亡抑制剂ferrostatin-1(Fer-1)干预后,MIN6细胞活力增加,Fe^(2+),MDA含量,ROS水平,ACSL4表达水平均显著降低,GSH,SOD含量明显升高(P<0.05)。GPX4以及通路蛋白SLC7A11、SLC3A2、GSS和GCLM表达水平显著升高(P<0.05)。结论GSPE能够通过上调Xc^(-)/GPX4信号通路拮抗MIN6细胞发生铁死亡。

Objective To investigate the protective effect and possible mechanism of grape seed proanthocyanidins(GSPE)on high-fat and high-sugar induced cell injury in mouse insulinoma MIN6 cells.Methods The cell viability was detected by CCK-8 method;intra-cellular ferrous ions(Fe^(2+))were detected by Lillie staining;glutathione(GSH),malondialdehyde(MDA)and superoxide dismutase(SOD)contents were detected;reactive oxygen species(ROS)levels were detected by fluorescent probe DCFH-DA labeling;western blot was used to detect iron death key proteins(GPX4 and ACSL4)and Xc^(-)/GPX4 signalling pathway proteins(SLC7A11,SLC3A2,GSS and GCLM)by western blot.Results High-fat and high-sugar treatment inhibited MIN6 cell viability,resulting in increased Fe^(2+),MDA content,ROS levels,decreased GSH,SOD content,increased ACSL4 expression levels and decreased GPX4 and pathway pro-teins SLC7A11,SLC3A2,GSS and GCLM expression levels.After the administration of different concentrations of GSPE or the ferrop-tosis inhibitor ferrostatin-1(Fer-1),MIN6 cell viability increased,Fe^(2+),MDA levels,ROS levels,ACSL4 expression levels were significantly reduced,and GSH and SOD levels were significantly increased(P<0.05).GPX4 and pathway proteins SLC7A11,SLC3A2,GSS and GCLM expression levels were significantly increased.SLC3A2,GSS and GCLM expression levels were significantly increased(P<0.05).Conclusion GSPE can antagonize ferroptosis in MIN6 cells through upregulation of the Xc^(-)/GPX4 signaling pathway.