雷帕霉素对高糖高脂诱导的MIN6细胞铁死亡的研究  

Rapamycin attenuates high glucose and high lipid-induced ferroptosis in MIN6 cells

作　　者：杨瑞瑞 王浩 陆恒 李海燕 王同玲 张丽媛 郭恒[1] 丁玉松[1] YANG Ruirui;WANG Hao;LU Heng;LI Haiyan;WANG Tongling;ZHANG Liyuan;GUO Heng;DING Yusong(Department of Preventive Medicine,School of Medicine,Shihezi University,Shihezi,Xinjiang 832000,China)

机构地区：[1]石河子大学医学院预防医学系,新疆石河子832000

出　　处：《石河子大学学报（自然科学版）》2023年第6期772-779,共8页Journal of Shihezi University(Natural Science)

基　　金：新疆生产建设兵团重点领域科技攻关计划项目(2021AB030)。

摘　　要：目的探讨自噬诱导剂能否通过激活自噬减少高糖高脂诱导的小鼠胰岛β细胞铁死亡。方法选取小鼠胰岛瘤MIN6细胞,通过25 mmol·L^(-1)高糖联合200μmol·L^(-1)棕榈酸钠(high Glucose and high Sodium Palmitate,GP)干预,建立胰岛β细胞损伤模型,在此基础上给予雷帕霉素(rapamycin,RAPA)及铁死亡诱导剂(erastin,Era)干预24 h后,用CCK8检测细胞活力;用试剂盒检测细胞亚铁离子、谷胱甘肽(glutathione,GSH)、超氧化物歧化酶(Superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)及活性氧(reactive oxygen species,ROS)水平;Western blot检测铁死亡相关蛋白谷胱甘肽过氧化物酶4(Glutathione Peroxidase 4,GPX4)和酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long-chain family member 4,ACSL4)、铁蛋白(Ferritin,FE)及自噬相关蛋白微管相关蛋白l轻链3Ⅱ(Microtubule-associated protein1 light chain3,LC3Ⅱ)、P62水平。结果与对照组相比,GP组及Era组MIN6细胞活力、GSH含量、SOD活力均降低,ROS、MDA及亚铁离子增多,GPX4蛋白表达水平降低,ACSL4、LC3Ⅱ、P62、FE蛋白表达升高(P<0.05)。与GP相比,RAPA干预后细胞活力增加,RAPA使亚铁离子、MDA及ROS水平降低,GSH、SOD含量、GPX4、LC3Ⅱ蛋白表达水平升高,ACSL4、P62、FE蛋白表达降低(P<0.05)。结论铁死亡参与GP诱导的MIN6细胞损伤,其机制可能与自噬流阻滞有关。雷帕霉素可改善β细胞铁死亡。Objective To investigate whether autophagy inducer can reduce ferroptosis of mouse pancreaticβ-cells induced by high-glucose and high-lipid through activation of autophagy.Methods The pancreaticβ-cell injury model was established by treating high glucose(25 mmol·L^(-1))and sodium palmitate(200μmol·L^(-1))(GP)in MIN6 cell.And on this basis,treated with rapamycin(RAPA)and erastin(Era).And the cell viability was detected by CCK8 assay.Furthermore,ferrous ion,the concentration of reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH)and the activity of superoxide dismutase(SOD)were detected by commercial kit.Glutathione Peroxidase 4(GPX4)and acyl-CoA synthetase long-chain family member 4(ACSL4),ferritin(Ferritin,FE)and Microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ)and P62 were detected by Western blot.Results GP and Era inhibited MIN6 cell viability,GSH content,SOD activity,increased ROS level,MDA and ferrous ion content,decreased GPX4 protein expression level and increased ACSL4,LC3II,P62,FE protein expression level(P<0.05).RAPA intervention increased cell viability,decreased ferrous ions,MDA and ROS levels,increased GSH,SOD content,increased GPX4,LC3Ⅱprotein expression,and decreased ACSL4,P62,and FE protein expression(P<0.05).Conclusion Ferroptosis is involved in GP-induced MIN6 cell damage,and the mechanism may be related to autophagic flux blockage.Rapamycin may improveβ-cell ferroptosis.

关 键 词：铁死亡 自噬 高糖高脂 胰岛Β细胞 

分 类 号：R151[医药卫生—营养与食品卫生学]